The property of emitting radiation while being irradiated. The radiation emitted is usually of longer wavelength than that incident or absorbed, e.g., a substance can be irradiated with invisible radiation and emit visible light. X-ray fluorescence is used in diagnosis.
Measurement of the intensity and quality of fluorescence.
Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye.
The use of instrumentation and techniques for visualizing material and details that cannot be seen by the unaided eye. It is usually done by enlarging images, transmitted by light or electron beams, with optical or magnetic lenses that magnify the entire image field. With scanning microscopy, images are generated by collecting output from the specimen in a point-by-point fashion, on a magnified scale, as it is scanned by a narrow beam of light or electrons, a laser, a conductive probe, or a topographical probe.
Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.
Microscopy in which the object is examined directly by an electron beam scanning the specimen point-by-point. The image is constructed by detecting the products of specimen interactions that are projected above the plane of the sample, such as backscattered electrons. Although SCANNING TRANSMISSION ELECTRON MICROSCOPY also scans the specimen point by point with the electron beam, the image is constructed by detecting the electrons, or their interaction products that are transmitted through the sample plane, so that is a form of TRANSMISSION ELECTRON MICROSCOPY.
A light microscopic technique in which only a small spot is illuminated and observed at a time. An image is constructed through point-by-point scanning of the field in this manner. Light sources may be conventional or laser, and fluorescence or transmitted observations are possible.
Measurement of the polarization of fluorescent light from solutions or microscopic specimens. It is used to provide information concerning molecular size, shape, and conformation, molecular anisotropy, electronic energy transfer, molecular interaction, including dye and coenzyme binding, and the antigen-antibody reaction.
A type of IN SITU HYBRIDIZATION in which target sequences are stained with fluorescent dye so their location and size can be determined using fluorescence microscopy. This staining is sufficiently distinct that the hybridization signal can be seen both in metaphase spreads and in interphase nuclei.
Agents that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags.
A type of FLUORESCENCE SPECTROSCOPY using two FLUORESCENT DYES with overlapping emission and absorption spectra, which is used to indicate proximity of labeled molecules. This technique is useful for studying interactions of molecules and PROTEIN FOLDING.
Fluorescence microscopy utilizing multiple low-energy photons to produce the excitation event of the fluorophore. Multiphoton microscopes have a simplified optical path in the emission side due to the lack of an emission pinhole, which is necessary with normal confocal microscopes. Ultimately this allows spatial isolation of the excitation event, enabling deeper imaging into optically thick tissue, while restricting photobleaching and phototoxicity to the area being imaged.
A type of scanning probe microscopy in which a probe systematically rides across the surface of a sample being scanned in a raster pattern. The vertical position is recorded as a spring attached to the probe rises and falls in response to peaks and valleys on the surface. These deflections produce a topographic map of the sample.
Electron microscopy in which the ELECTRONS or their reaction products that pass down through the specimen are imaged below the plane of the specimen.
Microscopy in which the samples are first stained immunocytochemically and then examined using an electron microscope. Immunoelectron microscopy is used extensively in diagnostic virology as part of very sensitive immunoassays.
A method used to study the lateral movement of MEMBRANE PROTEINS and LIPIDS. A small area of a cell membrane is bleached by laser light and the amount of time necessary for unbleached fluorescent marker-tagged proteins to diffuse back into the bleached site is a measurement of the cell membrane's fluidity. The diffusion coefficient of a protein or lipid in the membrane can be calculated from the data. (From Segen, Current Med Talk, 1995).
Protein analogs and derivatives of the Aequorea victoria green fluorescent protein that emit light (FLUORESCENCE) when excited with ULTRAVIOLET RAYS. They are used in REPORTER GENES in doing GENETIC TECHNIQUES. Numerous mutants have been made to emit other colors or be sensitive to pH.
Proteins which are involved in the phenomenon of light emission in living systems. Included are the "enzymatic" and "non-enzymatic" types of system with or without the presence of oxygen or co-factors.
Electron microscopy involving rapid freezing of the samples. The imaging of frozen-hydrated molecules and organelles permits the best possible resolution closest to the living state, free of chemical fixatives or stains.
Microscopy in which television cameras are used to brighten magnified images that are otherwise too dark to be seen with the naked eye. It is used frequently in TELEPATHOLOGY.
A family of 3,6-di(substituted-amino)-9-benzoate derivatives of xanthene that are used as dyes and as indicators for various metals; also used as fluorescent tracers in histochemistry.
The rate dynamics in chemical or physical systems.
A type of scanning probe microscopy in which a very sharp conducting needle is swept just a few angstroms above the surface of a sample. The tiny tunneling current that flows between the sample and the needle tip is measured, and from this are produced three-dimensional topographs. Due to the poor electron conductivity of most biological samples, thin metal coatings are deposited on the sample.
Microscopy using polarized light in which phenomena due to the preferential orientation of optical properties with respect to the vibration plane of the polarized light are made visible and correlated parameters are made measurable.
The science and application of a double-beam transmission interference microscope in which the illuminating light beam is split into two paths. One beam passes through the specimen while the other beam reflects off a reference mirror before joining and interfering with the other. The observed optical path difference between the two beams can be measured and used to discriminate minute differences in thickness and refraction of non-stained transparent specimens, such as living cells in culture.
An essential amino acid that is necessary for normal growth in infants and for NITROGEN balance in adults. It is a precursor of INDOLE ALKALOIDS in plants. It is a precursor of SEROTONIN (hence its use as an antidepressant and sleep aid). It can be a precursor to NIACIN, albeit inefficiently, in mammals.
The transfer of energy of a given form among different scales of motion. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed). It includes the transfer of kinetic energy and the transfer of chemical energy. The transfer of chemical energy from one molecule to another depends on proximity of molecules so it is often used as in techniques to measure distance such as the use of FORSTER RESONANCE ENERGY TRANSFER.
A family of spiro(isobenzofuran-1(3H),9'-(9H)xanthen)-3-one derivatives. These are used as dyes, as indicators for various metals, and as fluorescent labels in immunoassays.
Discrete concentrations of energy, apparently massless elementary particles, that move at the speed of light. They are the unit or quantum of electromagnetic radiation. Photons are emitted when electrons move from one energy state to another. (From Hawley's Condensed Chemical Dictionary, 11th ed)
Compounds that contain three methine groups. They are frequently used as cationic dyes used for differential staining of biological materials.
The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
A form of interference microscopy in which variations of the refracting index in the object are converted into variations of intensity in the image. This is achieved by the action of a phase plate.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
Light-induced change in a chromophore, resulting in the loss of its absorption of light of a particular wave length. The photon energy causes a conformational change in the photoreceptor proteins affecting PHOTOTRANSDUCTION. This occurs naturally in the retina (ADAPTATION, OCULAR) on long exposure to bright light. Photobleaching presents problems when occurring in PHOTODYNAMIC THERAPY, and in FLUORESCENCE MICROSCOPY. On the other hand, this phenomenon is exploited in the technique, FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING, allowing measurement of the movements of proteins and LIPIDS in the CELL MEMBRANE.
The use of light interaction (scattering, absorption, and fluorescence) with biological tissue to obtain morphologically based information. It includes measuring inherent tissue optical properties such as scattering, absorption, and autofluorescence; or optical properties of exogenous targeted fluorescent molecular probes such as those used in optical MOLECULAR IMAGING, or nontargeted optical CONTRAST AGENTS.
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
A type of TRANSMISSION ELECTRON MICROSCOPY in which the object is examined directly by an extremely narrow electron beam scanning the specimen point-by-point and using the reactions of the electrons that are transmitted through the specimen to create the image. It should not be confused with SCANNING ELECTRON MICROSCOPY.
Elements of limited time intervals, contributing to particular results or situations.
An analytical method for detecting and measuring FLUORESCENCE in compounds or targets such as cells, proteins, or nucleotides, or targets previously labeled with FLUORESCENCE AGENTS.
Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
An optical source that emits photons in a coherent beam. Light Amplification by Stimulated Emission of Radiation (LASER) is brought about using devices that transform light of varying frequencies into a single intense, nearly nondivergent beam of monochromatic radiation. Lasers operate in the infrared, visible, ultraviolet, or X-ray regions of the spectrum.
Fluorescent probe capable of being conjugated to tissue and proteins. It is used as a label in fluorescent antibody staining procedures as well as protein- and amino acid-binding techniques.
Scanning microscopy in which a very sharp probe is employed in close proximity to a surface, exploiting a particular surface-related property. When this property is local topography, the method is atomic force microscopy (MICROSCOPY, ATOMIC FORCE), and when it is local conductivity, the method is scanning tunneling microscopy (MICROSCOPY, SCANNING TUNNELING).
A phthalic indicator dye that appears yellow-green in normal tear film and bright green in a more alkaline medium such as the aqueous humor.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
Established cell cultures that have the potential to propagate indefinitely.
The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
A change from planar to elliptic polarization when an initially plane-polarized light wave traverses an optically active medium. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
The tendency of a gas or solute to pass from a point of higher pressure or concentration to a point of lower pressure or concentration and to distribute itself throughout the available space. Diffusion, especially FACILITATED DIFFUSION, is a major mechanism of BIOLOGICAL TRANSPORT.
The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.
A technique of inputting two-dimensional images into a computer and then enhancing or analyzing the imagery into a form that is more useful to the human observer.
A basic element found in nearly all organized tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
The use of molecularly targeted imaging probes to localize and/or monitor biochemical and cellular processes via various imaging modalities that include RADIONUCLIDE IMAGING; ULTRASONOGRAPHY; MAGNETIC RESONANCE IMAGING; FLUORESCENCE IMAGING; and MICROSCOPY.
A class of organic compounds which contain an anilino (phenylamino) group linked to a salt or ester of naphthalenesulfonic acid. They are frequently used as fluorescent dyes and sulfhydryl reagents.
Fluoroimmunoassay where detection of the hapten-antibody reaction is based on measurement of the increased polarization of fluorescence-labeled hapten when it is combined with antibody. The assay is very useful for the measurement of small haptenic antigens such as drugs at low concentrations.
A naphthalene derivative with carcinogenic action.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
That portion of the electromagnetic spectrum in the visible, ultraviolet, and infrared range.
The study of PHYSICAL PHENOMENA and PHYSICAL PROCESSES as applied to living things.
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
Artificial, single or multilaminar vesicles (made from lecithins or other lipids) that are used for the delivery of a variety of biological molecules or molecular complexes to cells, for example, drug delivery and gene transfer. They are also used to study membranes and membrane proteins.
Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.
Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.
Porphyrin derivatives containing magnesium that act to convert light energy in photosynthetic organisms.
The spectrometric analysis of fluorescent X-RAYS, i.e. X-rays emitted after bombarding matter with high energy particles such as PROTONS; ELECTRONS; or higher energy X-rays. Identification of ELEMENTS by this technique is based on the specific type of X-rays that are emitted which are characteristic of the specific elements in the material being analyzed. The characteristic X-rays are distinguished and/or quantified by either wavelength dispersive or energy dispersive methods.
Filamentous proteins that are the main constituent of the thin filaments of muscle fibers. The filaments (known also as filamentous or F-actin) can be dissociated into their globular subunits; each subunit is composed of a single polypeptide 375 amino acids long. This is known as globular or G-actin. In conjunction with MYOSINS, actin is responsible for the contraction and relaxation of muscle.
The physical characteristics and processes of biological systems.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)
Characteristics or attributes of the outer boundaries of objects, including molecules.
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
The part of a cell that contains the CYTOSOL and small structures excluding the CELL NUCLEUS; MITOCHONDRIA; and large VACUOLES. (Glick, Glossary of Biochemistry and Molecular Biology, 1990)
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment.
A colorless, odorless, highly water soluble vinyl monomer formed from the hydration of acrylonitrile. It is primarily used in research laboratories for electrophoresis, chromatography, and electron microscopy and in the sewage and wastewater treatment industries.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)
Compounds that contain a 1-dimethylaminonaphthalene-5-sulfonyl group.
A group of condensed ring hydrocarbons.
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
Layers of lipid molecules which are two molecules thick. Bilayer systems are frequently studied as models of biological membranes.
A scientific tool based on ULTRASONOGRAPHY and used not only for the observation of microstructure in metalwork but also in living tissue. In biomedical application, the acoustic propagation speed in normal and abnormal tissues can be quantified to distinguish their tissue elasticity and other properties.
Chemicals and substances that impart color including soluble dyes and insoluble pigments. They are used in INKS; PAINTS; and as INDICATORS AND REAGENTS.
A fluorescent compound that emits light only in specific configurations in certain lipid media. It is used as a tool in the study of membrane lipids.
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
Determination of the spectra of ultraviolet absorption by specific molecules in gases or liquids, for example Cl2, SO2, NO2, CS2, ozone, mercury vapor, and various unsaturated compounds. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
The development and use of techniques to study physical phenomena and construct structures in the nanoscale size range or smaller.
LIGHT, it's processes and properties, and the characteristics of materials interacting with it.
The process of generating three-dimensional images by electronic, photographic, or other methods. For example, three-dimensional images can be generated by assembling multiple tomographic images with the aid of a computer, while photographic 3-D images (HOLOGRAPHY) can be made by exposing film to the interference pattern created when two laser light sources shine on an object.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
A class of organic compounds that contains a naphthalene moiety linked to a sulfonic acid salt or ester.
Any of a variety of procedures which use biomolecular probes to measure the presence or concentration of biological molecules, biological structures, microorganisms, etc., by translating a biochemical interaction at the probe surface into a quantifiable physical signal.
The diversion of RADIATION (thermal, electromagnetic, or nuclear) from its original path as a result of interactions or collisions with atoms, molecules, or larger particles in the atmosphere or other media. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
Proteins found in any species of bacterium.
The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.
Derivatives of phosphatidic acids in which the phosphoric acid is bound in ester linkage to a choline moiety. Complete hydrolysis yields 1 mole of glycerol, phosphoric acid and choline and 2 moles of fatty acids.
Disruption of the non-covalent bonds and/or disulfide bonds responsible for maintaining the three-dimensional shape and activity of the native protein.
The motion of phospholipid molecules within the lipid bilayer, dependent on the classes of phospholipids present, their fatty acid composition and degree of unsaturation of the acyl chains, the cholesterol concentration, and temperature.
A specialized field of physics and engineering involved in studying the behavior and properties of light and the technology of analyzing, generating, transmitting, and manipulating ELECTROMAGNETIC RADIATION in the visible, infrared, and ultraviolet range.
Nanometer sized fragments of semiconductor crystalline material which emit PHOTONS. The wavelength is based on the quantum confinement size of the dot. They can be embedded in MICROBEADS for high throughput ANALYTICAL CHEMISTRY TECHNIQUES.
Proteins prepared by recombinant DNA technology.
Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.
Behavior of LIGHT and its interactions with itself and materials.
Relating to the size of solids.
A rigorously mathematical analysis of energy relationships (heat, work, temperature, and equilibrium). It describes systems whose states are determined by thermal parameters, such as temperature, in addition to mechanical and electromagnetic parameters. (From Hawley's Condensed Chemical Dictionary, 12th ed)
Nanometer-sized particles that are nanoscale in three dimensions. They include nanocrystaline materials; NANOCAPSULES; METAL NANOPARTICLES; DENDRIMERS, and QUANTUM DOTS. The uses of nanoparticles include DRUG DELIVERY SYSTEMS and cancer targeting and imaging.
Recording serial images of a process at regular intervals spaced out over a longer period of time than the time in which the recordings will be played back.
Substances used for the detection, identification, analysis, etc. of chemical, biological, or pathologic processes or conditions. Indicators are substances that change in physical appearance, e.g., color, at or approaching the endpoint of a chemical titration, e.g., on the passage between acidity and alkalinity. Reagents are substances used for the detection or determination of another substance by chemical or microscopical means, especially analysis. Types of reagents are precipitants, solvents, oxidizers, reducers, fluxes, and colorimetric reagents. (From Grant & Hackh's Chemical Dictionary, 5th ed, p301, p499)
The location of the atoms, groups or ions relative to one another in a molecule, as well as the number, type and location of covalent bonds.
A benzofuran derivative used as a protein reagent since the terminal N-NBD-protein conjugate possesses interesting fluorescence and spectral properties. It has also been used as a covalent inhibitor of both beef heart mitochondrial ATPase and bacterial ATPase.
The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.
The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.
A compound produced from succinyl-CoA and GLYCINE as an intermediate in heme synthesis. It is used as a PHOTOCHEMOTHERAPY for actinic KERATOSIS.
The deductive study of shape, quantity, and dependence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
A tricarbocyanine dye that is used diagnostically in liver function tests and to determine blood volume and cardiac output.
Compounds with three aromatic rings in linear arrangement with an OXYGEN in the center ring.
Compounds formed by the joining of smaller, usually repeating, units linked by covalent bonds. These compounds often form large macromolecules (e.g., BIOPOLYMERS; PLASTICS).
The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.
A technique encompassing morphometry, densitometry, neural networks, and expert systems that has numerous clinical and research applications and is particularly useful in anatomic pathology for the study of malignant lesions. The most common current application of image cytometry is for DNA analysis, followed by quantitation of immunohistochemical staining.
A cell line derived from cultured tumor cells.
Salts and esters of the 12-carbon saturated monocarboxylic acid--lauric acid.
A group of atoms or molecules attached to other molecules or cellular structures and used in studying the properties of these molecules and structures. Radioactive DNA or RNA sequences are used in MOLECULAR GENETICS to detect the presence of a complementary sequence by NUCLEIC ACID HYBRIDIZATION.
An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.
The level of protein structure in which regular hydrogen-bond interactions within contiguous stretches of polypeptide chain give rise to alpha helices, beta strands (which align to form beta sheets) or other types of coils. This is the first folding level of protein conformation.
Materials which have structured components with at least one dimension in the range of 1 to 100 nanometers. These include NANOCOMPOSITES; NANOPARTICLES; NANOTUBES; and NANOWIRES.
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
Methods of creating machines and devices.
The network of filaments, tubules, and interconnecting filamentous bridges which give shape, structure, and organization to the cytoplasm.
Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.
The characteristic three-dimensional shape of a molecule.
Slender, cylindrical filaments found in the cytoskeleton of plant and animal cells. They are composed of the protein TUBULIN and are influenced by TUBULIN MODULATORS.
Drugs that are pharmacologically inactive but when exposed to ultraviolet radiation or sunlight are converted to their active metabolite to produce a beneficial reaction affecting the diseased tissue. These compounds can be administered topically or systemically and have been used therapeutically to treat psoriasis and various types of neoplasms.
A yellow metallic element with the atomic symbol Au, atomic number 79, and atomic weight 197. It is used in jewelry, goldplating of other metals, as currency, and in dental restoration. Many of its clinical applications, such as ANTIRHEUMATIC AGENTS, are in the form of its salts.
Any visual display of structural or functional patterns of organs or tissues for diagnostic evaluation. It includes measuring physiologic and metabolic responses to physical and chemical stimuli, as well as ultramicroscopy.
Study of intracellular distribution of chemicals, reaction sites, enzymes, etc., by means of staining reactions, radioactive isotope uptake, selective metal distribution in electron microscopy, or other methods.
A strong organic base existing primarily as guanidium ions at physiological pH. It is found in the urine as a normal product of protein metabolism. It is also used in laboratory research as a protein denaturant. (From Martindale, the Extra Pharmacopoeia, 30th ed and Merck Index, 12th ed) It is also used in the treatment of myasthenia and as a fluorescent probe in HPLC.
Cellular uptake of extracellular materials within membrane-limited vacuoles or microvesicles. ENDOSOMES play a central role in endocytosis.
Lipids, predominantly phospholipids, cholesterol and small amounts of glycolipids found in membranes including cellular and intracellular membranes. These lipids may be arranged in bilayers in the membranes with integral proteins between the layers and peripheral proteins attached to the outside. Membrane lipids are required for active transport, several enzymatic activities and membrane formation.
Microscopy in which the image is formed by ultraviolet radiation and is displayed and recorded by means of photographic film.
The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.
Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).
The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.
A trypanocidal agent and possible antiviral agent that is widely used in experimental cell biology and biochemistry. Ethidium has several experimentally useful properties including binding to nucleic acids, noncompetitive inhibition of nicotinic acetylcholine receptors, and fluorescence among others. It is most commonly used as the bromide.
A cationic cytochemical stain specific for cell nuclei, especially DNA. It is used as a supravital stain and in fluorescence cytochemistry. It may cause mutations in microorganisms.
A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
The use of devices which use detector molecules to detect, investigate, or analyze other molecules, macromolecules, molecular aggregates, or organisms.
Artificially produced membranes, such as semipermeable membranes used in artificial kidney dialysis (RENAL DIALYSIS), monomolecular and bimolecular membranes used as models to simulate biological CELL MEMBRANES. These membranes are also used in the process of GUIDED TISSUE REGENERATION.
A form of fluorescent antibody technique commonly used to detect serum antibodies and immune complexes in tissues and microorganisms in specimens from patients with infectious diseases. The technique involves formation of an antigen-antibody complex which is labeled with fluorescein-conjugated anti-immunoglobulin antibody. (From Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984)
A group of glucose polymers made by certain bacteria. Dextrans are used therapeutically as plasma volume expanders and anticoagulants. They are also commonly used in biological experimentation and in industry for a wide variety of purposes.
Silver. An element with the atomic symbol Ag, atomic number 47, and atomic weight 107.87. It is a soft metal that is used medically in surgical instruments, dental prostheses, and alloys. Long-continued use of silver salts can lead to a form of poisoning known as ARGYRIA.
A clear, odorless, tasteless liquid that is essential for most animal and plant life and is an excellent solvent for many substances. The chemical formula is hydrogen oxide (H2O). (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
A broad class of substances containing carbon and its derivatives. Many of these chemicals will frequently contain hydrogen with or without oxygen, nitrogen, sulfur, phosphorus, and other elements. They exist in either carbon chain or carbon ring form.
Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.
Porphyrins with four methyl, two vinyl, and two propionic acid side chains attached to the pyrrole rings. Protoporphyrin IX occurs in hemoglobin, myoglobin, and most of the cytochromes.
Nanoparticles produced from metals whose uses include biosensors, optics, and catalysts. In biomedical applications the particles frequently involve the noble metals, especially gold and silver.
A noninvasive technique that uses the differential absorption properties of hemoglobin and myoglobin to evaluate tissue oxygenation and indirectly can measure regional hemodynamics and blood flow. Near-infrared light (NIR) can propagate through tissues and at particular wavelengths is differentially absorbed by oxygenated vs. deoxygenated forms of hemoglobin and myoglobin. Illumination of intact tissue with NIR allows qualitative assessment of changes in the tissue concentration of these molecules. The analysis is also used to determine body composition.
Preparation for electron microscopy of minute replicas of exposed surfaces of the cell which have been ruptured in the frozen state. The specimen is frozen, then cleaved under high vacuum at the same temperature. The exposed surface is shadowed with carbon and platinum and coated with carbon to obtain a carbon replica.
That portion of the electromagnetic spectrum usually sensed as heat. Infrared wavelengths are longer than those of visible light, extending into the microwave frequencies. They are used therapeutically as heat, and also to warm food in restaurants.
Determination, by measurement or comparison with a standard, of the correct value of each scale reading on a meter or other measuring instrument; or determination of the settings of a control device that correspond to particular values of voltage, current, frequency or other output.
Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed)
The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability.
Analysis of the intensity of Raman scattering of monochromatic light as a function of frequency of the scattered light.
The assembly of the QUATERNARY PROTEIN STRUCTURE of multimeric proteins (MULTIPROTEIN COMPLEXES) from their composite PROTEIN SUBUNITS.
Colorless, odorless crystals that are used extensively in research laboratories for the preparation of polyacrylamide gels for electrophoresis and in organic synthesis, and polymerization. Some of its polymers are used in sewage and wastewater treatment, permanent press fabrics, and as soil conditioning agents.
Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.
The evaluation of incidents involving the loss of function of a device. These evaluations are used for a variety of purposes such as to determine the failure rates, the causes of failures, costs of failures, and the reliability and maintainability of devices.
Antibodies produced by a single clone of cells.
The technique of using a cryostat or freezing microtome, in which the temperature is regulated to -20 degrees Celsius, to cut ultrathin frozen sections for microscopic (usually, electron microscopic) examination.
Small uniformly-sized spherical particles, of micrometer dimensions, frequently labeled with radioisotopes or various reagents acting as tags or markers.
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system.
The process by which two molecules of the same chemical composition form a condensation product or polymer.
Pieces of glass or other transparent materials used for magnification or increased visual acuity.
Methods used to study CELLS.
Projection of near-IR light (INFRARED RAYS), in the 700-1000 nm region, across an object in parallel beams to an array of sensitive photodetectors. This is repeated at various angles and a mathematical reconstruction provides three dimensional MEDICAL IMAGING of tissues. Based on the relative transparency of tissues to this spectra, it has been used to monitor local oxygenation, brain and joints.
The measurement of the amplitude of the components of a complex waveform throughout the frequency range of the waveform. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
Transport proteins that carry specific substances in the blood or across cell membranes.
Fibers composed of MICROFILAMENT PROTEINS, which are predominately ACTIN. They are the smallest of the cytoskeletal filaments.
Particles consisting of aggregates of molecules held loosely together by secondary bonds. The surface of micelles are usually comprised of amphiphatic compounds that are oriented in a way that minimizes the energy of interaction between the micelle and its environment. Liquids that contain large numbers of suspended micelles are referred to as EMULSIONS.
A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).
Inorganic or organic compounds that contain boron as an integral part of the molecule.
The visually perceived property of objects created by absorption or reflection of specific wavelengths of light.
The technique of using a microtome to cut thin or ultrathin sections of tissues embedded in a supporting substance. The microtome is an instrument that hold a steel, glass or diamond knife in clamps at an angle to the blocks of prepared tissues, which it cuts in sections of equal thickness.
Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.
The relationship between the dose of an administered drug and the response of the organism to the drug.
Europium. An element of the rare earth family of metals. It has the atomic symbol Eu, atomic number 63, and atomic weight 152. Europium is used in the form of its salts as coatings for cathode ray tubes and in the form of its organic derivatives as shift reagents in NMR spectroscopy.
The resistance that a gaseous or liquid system offers to flow when it is subjected to shear stress. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed)
Synthetic phospholipid used in liposomes and lipid bilayers to study biological membranes. It is also a major constituent of PULMONARY SURFACTANTS.
The homogeneous mixtures formed by the mixing of a solid, liquid, or gaseous substance (solute) with a liquid (the solvent), from which the dissolved substances can be recovered by physical processes. (From Grant & Hackh's Chemical Dictionary, 5th ed)

Association of snRNA genes with coiled bodies is mediated by nascent snRNA transcripts. (1/23915)

BACKGROUND: Coiled bodies are nuclear organelles that are highly enriched in small nuclear ribonucleoproteins (snRNPs) and certain basal transcription factors. Surprisingly, coiled bodies not only contain mature U snRNPs but also associate with specific chromosomal loci, including gene clusters that encode U snRNAs and histone messenger RNAs. The mechanism(s) by which coiled bodies associate with these genes is completely unknown. RESULTS: Using stable cell lines, we show that artificial tandem arrays of human U1 and U2 snRNA genes colocalize with coiled bodies and that the frequency of the colocalization depends directly on the transcriptional activity of the array. Association of the genes with coiled bodies was abolished when the artificial U2 arrays contained promoter mutations that prevent transcription or when RNA polymerase II transcription was globally inhibited by alpha-amanitin. Remarkably, the association was also abolished when the U2 snRNA coding regions were replaced by heterologous sequences. CONCLUSIONS: The requirement for the U2 snRNA coding region indicates that association of snRNA genes with coiled bodies is mediated by the nascent U2 RNA itself, not by DNA or DNA-bound proteins. Our data provide the first evidence that association of genes with a nuclear organelle can be directed by an RNA and suggest an autogenous feedback regulation model.  (+info)

Deletion analysis of the Drosophila Inscuteable protein reveals domains for cortical localization and asymmetric localization. (2/23915)

The Drosophila Inscuteable protein acts as a key regulator of asymmetric cell division during the development of the nervous system [1] [2]. In neuroblasts, Inscuteable localizes into an apical cortical crescent during late interphase and most of mitosis. During mitosis, Inscuteable is required for the correct apical-basal orientation of the mitotic spindle and for the asymmetric segregation of the proteins Numb [3] [4] [5], Prospero [5] [6] [7] and Miranda [8] [9] into the basal daughter cell. When Inscuteable is ectopically expressed in epidermal cells, which normally orient their mitotic spindle parallel to the embryo surface, these cells reorient their mitotic spindle and divide perpendicularly to the surface [1]. Like the Inscuteable protein, the inscuteable RNA is asymmetrically localized [10]. We show here that inscuteable RNA localization is not required for Inscuteable protein localization. We found that a central 364 amino acid domain - the Inscuteable asymmetry domain - was necessary and sufficient for Inscuteable localization and function. Within this domain, a separate 100 amino acid region was required for asymmetric localization along the cortex, whereas a 158 amino acid region directed localization to the cell cortex. The same 158 amino acid fragment could localize asymmetrically when coexpressed with the full-length protein, however, and could bind to Inscuteable in vitro, suggesting that this domain may be involved in the self-association of Inscuteable in vivo.  (+info)

Evidence for F-actin-dependent and -independent mechanisms involved in assembly and stability of the medial actomyosin ring in fission yeast. (3/23915)

Cell division in a number of eukaryotes, including the fission yeast Schizosaccharomyces pombe, is achieved through a medially placed actomyosin-based contractile ring. Although several components of the actomyosin ring have been identified, the mechanisms regulating ring assembly are still not understood. Here, we show by biochemical and mutational studies that the S.pombe actomyosin ring component Cdc4p is a light chain associated with Myo2p, a myosin II heavy chain. Localization of Myo2p to the medial ring depended on Cdc4p function, whereas localization of Cdc4p at the division site was independent of Myo2p. Interestingly, the actin-binding and motor domains of Myo2p are not required for its accumulation at the division site although the motor activity of Myo2p is essential for assembly of a normal actomyosin ring. The initial assembly of Myo2p and Cdc4p at the division site requires a functional F-actin cytoskeleton. Once established, however, F-actin is not required for the maintenance of Cdc4p and Myo2p medial rings, suggesting that the attachment of Cdc4p and Myo2p to the division site involves proteins other than actin itself.  (+info)

Deletion of a region that is a candidate for the difference between the deletion forms of hereditary persistence of fetal hemoglobin and deltabeta-thalassemia affects beta- but not gamma-globin gene expression. (4/23915)

The analysis of a number of cases of beta-globin thalassemia and hereditary persistence of fetal hemoglobin (HPFH) due to large deletions in the beta-globin locus has led to the identification of several DNA elements that have been implicated in the switch from human fetal gamma- to adult beta-globin gene expression. We have tested this hypothesis for an element that covers the minimal distance between the thalassemia and HPFH deletions and is thought to be responsible for the difference between a deletion HPFH and deltabeta-thalassemia, located 5' of the delta-globin gene. This element has been deleted from a yeast artificial chromosome (YAC) containing the complete human beta-globin locus. Analysis of this modified YAC in transgenic mice shows that early embryonic expression is unaffected, but in the fetal liver it is subject to position effects. In addition, the efficiency of transcription of the beta-globin gene is decreased, but the developmental silencing of the gamma-globin genes is unaffected by the deletion. These results show that the deleted element is involved in the activation of the beta-globin gene perhaps through the loss of a structural function required for gene activation by long-range interactions.  (+info)

The disulfide-bonded loop of chromogranin B mediates membrane binding and directs sorting from the trans-Golgi network to secretory granules. (5/23915)

The disulfide-bonded loop of chromogranin B (CgB), a regulated secretory protein with widespread distribution in neuroendocrine cells, is known to be essential for the sorting of CgB from the trans-Golgi network (TGN) to immature secretory granules. Here we show that this loop, when fused to the constitutively secreted protein alpha1-antitrypsin (AT), is sufficient to direct the fusion protein to secretory granules. Importantly, the sorting efficiency of the AT reporter protein bearing two loops (E2/3-AT-E2/3) is much higher compared with that of AT with a single disulfide-bonded loop. In contrast to endogenous CgB, E2/3-AT-E2/3 does not undergo Ca2+/pH-dependent aggregation in the TGN. Furthermore, the disulfide-bonded loop of CgB mediates membrane binding in the TGN and does so with 5-fold higher efficiency if two loops are present on the reporter protein. The latter finding supports the concept that under physiological conditions, aggregates of CgB are the sorted units of cargo which have multiple loops on their surface leading to high membrane binding and sorting efficiency of CgB in the TGN.  (+info)

Optical mapping of Plasmodium falciparum chromosome 2. (6/23915)

Detailed restriction maps of microbial genomes are a valuable resource in genome sequencing studies but are toilsome to construct by contig construction of maps derived from cloned DNA. Analysis of genomic DNA enables large stretches of the genome to be mapped and circumvents library construction and associated cloning artifacts. We used pulsed-field gel electrophoresis purified Plasmodium falciparum chromosome 2 DNA as the starting material for optical mapping, a system for making ordered restriction maps from ensembles of individual DNA molecules. DNA molecules were bound to derivatized glass surfaces, cleaved with NheI or BamHI, and imaged by digital fluorescence microscopy. Large pieces of the chromosome containing ordered DNA restriction fragments were mapped. Maps were assembled from 50 molecules producing an average contig depth of 15 molecules and high-resolution restriction maps covering the entire chromosome. Chromosome 2 was found to be 976 kb by optical mapping with NheI, and 946 kb with BamHI, which compares closely to the published size of 947 kb from large-scale sequencing. The maps were used to further verify assemblies from the plasmid library used for sequencing. Maps generated in silico from the sequence data were compared to the optical mapping data, and good correspondence was found. Such high-resolution restriction maps may become an indispensable resource for large-scale genome sequencing projects.  (+info)

Inducible long-term gene expression in brain with adeno-associated virus gene transfer. (7/23915)

Recombinant adeno-associated virus (rAAV) vectors hold promise for treating a number of neurological disorders due to the ability to deliver long-term gene expression without toxicity or immune response. Critical to these endeavors will be controlled expression of the therapeutic gene in target cells. We have constructed and tested a dual cassette rAAV vector carrying a reporter gene under the control of the tetracycline-responsive system and the tetracycline transactivator. Transduction in vitro resulted in stable expression from the vector that can be suppressed 20-fold by tetracycline treatment. In vivo experiments, carried out to 6 weeks, demonstrated that vector-transduced expression is sustained until doxycycline administration upon which reporter gene expression is reduced. Moreover, the suppression of vector-driven expression can be reversed by removal of the drug. These studies demonstrate long-term regulated gene expression from rAAV vectors. This system will provide a valuable approach for controlling vector gene expression both in vitro and in vivo.  (+info)

Transcriptional regulation and induction of apoptosis: implications for the use of monomeric p53 variants in gene therapy. (8/23915)

The p53 tumour suppressor protein is a transcriptional activator, which can induce cell cycle arrest and apoptosis. p53 Gene mutations occur in more than 50% of all human tumours. Reintroduction of wild-type p53 but also of oligomerisation-independent p53 variants into tumour cells by gene transfer methods has been considered. We have investigated the biological properties of two carboxy-terminal deletion mutants of p53, p53 delta 300 (comprising amino acids 1-300) and p53 delta 326 (amino acids 1-326), to evaluate their potential deployment in gene therapy. Transactivation was measured in transiently transfected HeLa and SKBR3 cells. Both monomeric variants showed reduced activities compared with wild-type p53. Individual promoters were differently affected. In contrast to wild-type p53, monomeric variants were not able to induce apoptosis. We also provided wild-type p53 and p53 delta 326 with tetracycline-regulated promoters and stably introduced these constructs into Saos2 and SKBR3 cells. Upon induction, wild-type p53 expressing cells, but not p53 delta 326 expressing cells underwent apoptosis. Consistently, only wild-type p53 expressing cells accumulated p21/waf1/cip1 mRNA and protein and showed increased bax, Gadd45 and mdm2 mRNA. Neither wild-type p53 nor p53 delta 326 repressed the transcription of the IGF-1R gene in these cell lines. We conclude that the transactivation potential of monomeric, carboxy-terminally truncated p53 is not sufficient to cause induction of the endogenous target genes which trigger apoptosis.  (+info)

NIEF manager, Dr. Otero, participated in the Quantitative Fluorescence Microscopy course held at the MDI Biological Laboratory in Bar Harbor, ME. This international course in fluorescence microscopy and imaging provided the opportunity to train in several high-end microscopes and imaging analysis software and take lectures that cover the theory, mechanics, and applications of this biomedical technology.. Dr. Otero received a competitive scholarship from MDI Biological Laboratory Education and won the Chroma Cube Award for providing an outstanding individual scientific presentation at the course.. #gallery-1{margin:auto}#gallery-1 .gallery-item{float:left;margin-top:10px;text-align:center;width:33%}#gallery-1 img{border:2px solid #cfcfcf}#gallery-1 .gallery-caption{margin-left:0} ...
Total Internal Reflection Microscopy (TIRM) is a sensitive non-invasive technique to measure the interaction potentials between a colloidal particle and a wall with femtonewton resolution. The equilibrium distribution of the particle-wall separation distance z is sampled monitoring the intensity I scattered by the Brownian particle under evanescent illumination. Central to the data analysis is the knowledge of the relation between I and the corresponding z, which typically must be known a priori. This poses considerable constraints to the experimental conditions where TIRM can be applied (short penetration depth of the evanescent wave, transparent surfaces). Here, we introduce a method to experimentally determine I(z) by relying only on the distance-dependent particle-wall hydrodynamic interactions. We demonstrate that this method largely extends the range of conditions accessible with TIRM, and even allows measurements on highly reflecting gold surfaces where multiple reflections lead to a ...
Total internal reflection microscopy (TIRM) is a sensitive non-invasive technique to measure the interaction potentials between a colloidal particle and a wall with femtonewton resolution. Our work has largely extended the range of conditions accessible with TIRM, e.g. gold coated surfaces or large penetration depths.
Imaging single-channel Ca21 signals by total internal reflection fluorescence microscopy. (A) Schematic of the TIRFM imaging system. The 488-nm beam from an argon ion laser (50 mW) passes through a 53 beam expander (BE) and is focused by a lens (FL; f ¼ 150 mm) via a dichroic mirror (DM)to a spot at the back focal plane of the microscope objective lens (Olympus TIRFM 603, oil immersion, NA ¼ 1.45). The focusing lens is mounted on a micrometer-driven translation stage, so that the laser beam can be adjusted to enter the periphery of the objective aperture so as to achieve total internal reflection at the interface between the cover glass and the aqueous bathing medium. An adjustable rectangular knife-blade aperture (A) located at a conjugate image plane defines the field of excitation. Fluorescence excited in the specimen by the evanescent wave is collected by the same objective, passes through the dichroic mirror and a barrier filter (BF) blocking the laser wavelength, and is imaged by an ...
Total internal reflection fluorescence microscopy (TIRFM) is an elegant optical technique utilized to observe single molecule fluorescence at surfaces and interfaces. This section is an index to our discussions, references, and interactive Java tutorials that describe TIRFM.
The regulated trafficking or exocytosis of cargo-containing vesicles to the cell surface is fundamental to all cells. By coupling the technology of fluorescently tagged fusion proteins with total internal reflection fluorescence microscopy (TIRFM), it is possible to achieve the high spatio-temporal resolution required to study the dynamics of sub-plasma membrane vesicle trafficking and exocytosis. TIRFM has been used in a number of cell types to visualize and dissect the various steps of exocytosis revealing how molecules identified via genetic and/or biochemical approaches are involved in the regulation of this process. Here, we summarize the contribution of TIRFM to our understanding of the mechanism of exocytosis and discuss the novel methods of analysis that are required to exploit the large volumes of data that can be produced using this technique.
We report the use of a high-refractive-index aplanatic solid immersion lens (ASIL) in total internal reflection fluorescence (TIRF) microscopy. This new solid immersion total internal reflection fluorescence (SITIRF) microscopy allows highly confined surface imaging with a significantly reduced imaging depth compared with conventional TIRF microscopy. We explore the application of a high refractive index, low optical dispersion material zirconium dioxide in the SITIRF microscope and also introduce a novel system design which enables the SITIRF microscope to work either in the epi-fluorescence or TIRF modes with variable illumination angles. We use both synthetic and biological samples to demonstrate that the imaging depth in the SITIRF microscope can be confined to a few tens of nanometers. SITIRF microscopy has the advantages of performing highly selective imaging and high-resolution high-contrast imaging. Potential applications in biological imaging and future developments of SITIRF microscopy ...
TY - JOUR. T1 - Real time imaging of single fluorophores on moving actin with an epifluorescence microscope. AU - Sase, I.. AU - Miyata, H.. AU - Corrie, J. E T. AU - Craik, J. S.. AU - Kinosita, K.. PY - 1995. Y1 - 1995. N2 - Relatively simple modifications of an ordinary epifluorescence microscope have greatly reduced its background luminescence, allowing continuous and real time imaging of single fluorophores in an aqueous medium. Main modifications were changing the excitation light path and setting an aperture stop so that stray light does not scatter inside the microscope. A simple and accurate method using actin filaments is presented to establish the singularity of the observed fluorophores. It was possible, at the video rate of 30 frames/s, to image individual tetramethylrhodamine fluorophores bound to actin filaments sliding over heavy meromyosin. The successful imaging of moving fluorophores demonstrates that conventional microscopes may become a routine tool for studying dynamic ...
Anytime: Self-serve breakfast in the dining hall. 09:00-09:30 Introduction Welcome, Introductions, Define Course Goals (Piston and St. Croix). 09:30-10:30 The Microscope 1 Advances in Modern Imaging for Biological Problems (Kenworthy). 10:30-11:00 Break at Maren Conference Center. 11:00-12:00 The Microscope 2 Transmitted Light Contrast in Microscopy (Piston). 12:00-13:00 Lunch. 13:00-14:45 LAB 1 TRANSMITTED LIGHT MICROSCOPY. 15:00-15:45 The Microscope 3 Fluorescence Microscopy and Quantitation (Piston). 15:45-16:45 Live Cell Imaging 1 Green-Fluorescent Protein (Kremers). 16:45-17:00 Break. 17:00-18:00 Digital Imaging 1 Digital Imaging, Resolution, and Detectors (Watkins). 18:00-19:00 Dinner. 19:00-20:00 Digital Imaging 2 Optical Filters and Brightness Demonstration (Watkins et al.). 20:30-23:00 LAB 2 STANDARD FLUORESCENCE MICROSCOPY. ...
Friday, May 16, 2014. 16:00 Arrival on campus, check-in to housing, get keys at Maren Conference Center. 18:00 Dinner and Social at Co-Op. 19:30 Reception at Maren Conference Center and conference material pickup. Late arrival keys placed in Dining Hall keybox for pick-up. Saturday, May 17, 2014. anytime self-serve breakfast in the dining hall. 09:00-09:30 Introduction Welcome, Introductions, Define Course Goals (Piston and St. Croix). 09:30-10:30 The Microscope 1 Advances in Modern Imaging for Biological Problems (Kenworthy). 10:30-11:00 Break at Maren Conference Center. 11:00-12:00 The Microscope 2 Transmitted Light Contrast in Microscopy (Piston). 12:00-13:00 Lunch. 13:00-14:45 LAB 1 TRANSMITTED LIGHT MICROSCOPY. 15:00-15:45 The Microscope 3 Fluorescence Microscopy and Quantitation (Piston). 15:45-16:45 Live Cell Imaging 1 Green-Fluorescent Protein (Kremers). 16:45-17:00 Break. 17:00-18:00 Digital Imaging 1 Digital Imaging, Resolution, and Detectors (Watkins). 18:00-19:00 Dinner. 19:00-20:00 ...
In 2008, the first international Theodor Förster lecture series took place at the University of Cambridge http://laser.ceb.cam.ac.uk/foerster. Throughout the year, leading researchers were invited from all over the world to hold talks and engage with the life science community at Cambridge. The focus was on developments in quantitative optical microscopy techniques, which are revolutionizing research in the biological sciences today.. The name of the series commemorates the brilliant contributions of Theodor Förster, who, 60 years ago, published his seminal paper on the quantitative theory of electronic energy transfer (Förster 1948). The process, now known as Förster resonance energy transfer (FRET), takes place frequently in nature and refers to the non-radiative transport of energy from a donor to an acceptor molecule. Förster recognized that through a sequence of such interactions, energy can migrate over distances much larger than the molecular scale and in one sweep he had resolved ...
In electromagnetics, an evanescent field, or evanescent wave, is an oscillating electric and/or magnetic field that does not propagate as an electromagnetic wave but whose energy is spatially concentrated in the vicinity of the source (oscillating charges and currents). Even when there in fact is an electromagnetic wave produced (e.g., by a transmitting antenna) one can still identify as an evanescent field the component of the electric or magnetic field that cannot be attributed to the propagating wave observed at a distance of many wavelengths (such as the far field of a transmitting antenna). A hallmark of an evanescent field is that there is no net energy flow in that region. Since the net flow of electromagnetic energy is given by the average Poynting vector, that means that the Poynting vector in these regions, as averaged over a complete oscillation cycle, is zero. In many cases one cannot simply say that a field is or is not evanescent. For instance, in the above illustration energy is ...
Topology of cell adhesion on a substrate is measured with nanometre precision by variable-angle total internal reflection fluorescence...
Wide-field fluorescence microscopy is a widely applied imaging technique used to examine cells and investigate their internal structures.
We describe a method of obtaining optical sectioning with a standard wide-field fluorescence microscope. The method involves acquiring two images, one with nonuniform illumination (in our case, speckle) and another with uniform illumination (in our case, randomized speckle). An evaluation of the loc …
Principal Investigator:FUNATSU Takashi, Project Period (FY):2009-07-23 - 2014-03-31, Research Category:Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Project Area:Integrative understanding of biological processes mediated by transient macromolecular complexes; New technology for visualizing physiologically metastable states.
A wide range of optical designs for TIRFM have been employed with both inverted and upright microscope configurations. This discussion focuses on TIRFM configurations that utilize a prism to direct light toward the TIR interface.
Basic molecular mechanisms of cell surface receptors that mediate transmembrane signals can be elucidated by integrating information from multiple interdisciplinary approaches. Our studies focus on the receptor (FceRI) for immunoglobulin E (IgE) that plays a central role in the allergic response and serves as a model for other types of immune receptors. Binding and cross-linking of IgE-FceRI complexes by antigen initiates signal transduction resulting in cell activation and release of chemical mediators.. We measure kinetics and thermodynamics of binding and cross-linking between cell-bound IgE and structurally defined ligands with fluorescence methods and analyze with realistic theoretical models to determine features that are critical for signaling. We employ quantitative fluorescence microscopy, including confocal imaging and total internal reflection fluorescence (TIRF) microscopy, to monitor changes in the distribution and dynamics of the receptor and signaling components (and genetically ...
Single molecule fluorescence measurements are used to probe the effects of GM1 in DPPC monolayers. Langmuir-Blodgett films of GM1 and DPPC were doped with ~10−8 mol% of the fluorescent lipid probe, BODIPY-PC, and transferred onto glass substrates at 23 mN/m. As shown previously, the individual orientation of each BODIPY-PC probe in the membrane can be measured using defocused polarized total internal reflection fluorescence microscopy, revealing changes in film properties at the molecular level. Here, BODIPY-PC tilt angle histograms are used to characterize the effects of GM1 in DPPC films from 0.05 mol% to 100 mol% GM1. At high GM1 levels (,5 mol% GM1), trends in the single molecule measurements agree with previous bulk measurements showing the turnover from condensing to expanding influence of GM1 at ~20 mol%, thus validating the single molecule approach. At biologically relevant, low concentrations of GM1 (,5 mol% GM1), where bulk fluorescence measurements are less informative, the single ...
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BILLERICA, Mass. -- Bruker today announced that it has acquired Vutara Incorporated, a technology leader in high-speed, three-dimensional (3D), ...
As a high-resolution wide-field near-surface microscopy, total internal reflection fluorescence microscopy (TIRFM) has been widely applied for the study of biomolecules. Unlike those costly, sample consuming and time consuming traditional detection assays, the application of TIRFM enable the direct quantification of biomolecules in a sample pretreatment and enrichment free fashion. Taking advantages of the TIRFM imaging system, in this thesis we have applied the TIRFM imaging system to directly quantify the content of different cancer associated biomarkers. Four different detection approaches for direct cancer biomarkers quantification with the aid of TIRFM were herein presented respectively. In Chapter 2, a direct quantification of nasopharyngeal carcinoma associated miRNAs was described. In the assay, five different miRNAs were chosen as the target analytes, which hybridized with the synthetic complementary LNA, probe in solution. The duplex was labeled with intercalating fluorescence dye YOYO-1 and
Total internal reflection fluorescence (TIRF) microscopy is a powerful technique. It provides extremely thin axial sectioning with excellent signal-to
Total internal reflection fluorescence (TIRF) microscopy is a high‐contrast imaging technique suitable for observing biological events that occur on or near the cell membrane
TY - JOUR. T1 - Threshold-free evaluation of near-surface diffusion and adsorption-dominated motion from single-molecule tracking data of single-stranded DNA through total internal reflection fluorescence microscopy. AU - Hanasaki, Itsuo. AU - Uehara, Satoshi. AU - Arai, Yoshiyuki. AU - Nagai, Takeharu. AU - Kawano, Satoyuki. PY - 2015/12. Y1 - 2015/12. N2 - Total internal reflection fluorescence (TIRF) microscopy enables the single-molecule observation in liquid near substrate surface. However, the evaluation of the diffusion from their individually-tracked positions entails the difficulty in the treatment of molecular adsorption on the substrate. We propose a novel technique to evaluate them, and two types of near-surface Brownian motion were determined for DNA. One is the diffusion near glass surface, and the other is the adsorption-dominated motion, which is also found to be diffusive rather than anchored to the substrate. Our technique does not require the threshold values for the ...
Fluorescence microscopy images of cells double stained with phalloidin for actin filaments (red) and DAPI for nuclei (blue) on Ti (A) and USP-Ti (B) samples.Abb
Video articles in JoVE about tirf include A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors, SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy, Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy, A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins, Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy, Single Molecule Fluorescence Microscopy on Planar Supported Bilayers, Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking, Imaging Cell Membrane Injury and Subcellular Processes Involved in Repair, Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes, Counting Proteins in Single Cells with Addressable
Dr. Jorge Bernardino de la Serna (JbdlS) is a Senior Lecturer at the National Heart and Lung Institute, Imperial College London, and Visiting Scientist at the United Kingdom Research and Innovation. JBdlS has a markedly multidisciplinary research track record; he has ~25 publications in the last 5 years. JBdlSs research has been highlighted on 6 journal covers for his work on protein-lipid localisation and dynamics; Tcell lipid-protein interactions in atopic dermatitis; advanced quantitative fluorescence microscopy and spectroscopy; and peptide/peptoid-protein interactions with biological films. All these publications had in common the employment of highly-advanced microscopy techniques. JBdlS last publication in ACSnano were he bserved the theragnostic effect in Gram Negative Bacteria of a fluorescent molecule at the nanoscale. This article has been highlighted in The Telegraph and BBC Sheffield. JBdlS has a track record of microscopy method development and customisation to understand ...
Dr. Jorge Bernardino de la Serna (JbdlS) is a Senior Lecturer at the National Heart and Lung Institute, Imperial College London, and Visiting Scientist at the United Kingdom Research and Innovation. JBdlS has a markedly multidisciplinary research track record; he has ~25 publications in the last 5 years. JBdlSs research has been highlighted on 6 journal covers for his work on protein-lipid localisation and dynamics; Tcell lipid-protein interactions in atopic dermatitis; advanced quantitative fluorescence microscopy and spectroscopy; and peptide/peptoid-protein interactions with biological films. All these publications had in common the employment of highly-advanced microscopy techniques. JBdlS last publication in ACSnano were he bserved the theragnostic effect in Gram Negative Bacteria of a fluorescent molecule at the nanoscale. This article has been highlighted in The Telegraph and BBC Sheffield. JBdlS has a track record of microscopy method development and customisation to understand ...
In this study we show that as a single agent in Hct116 cells in vitro, SN-38 induces cell cycle arrest without cell death. This correlates to the absence of CRs observed in vivo with CPT-11 alone. The addition of flavopiridol to SN-38-treated Hct116 cells caused cell death in vitro, and in vivo this translated into greater tumor regression as well as CRs. With clonogenic assays, we observed that SN-38-treated cells remained attached to the plate as viable single cells. These cells did not form colonies because cell cycle arrest by SN-38, and this resulted in the inhibition of colony formation. However, SN-38 alone did not induce apoptosis; therefore, we did not observe PARP cleavage or caspase-3 activation. In contrast, flavopiridol added to SN-38-treated Hct116 cells induced significant apoptosis as indicated by quantitative fluorescence microscopy assays, PARP cleavage, and caspase-3 activation. We observed neither single viable cells nor colonies by clonogenic assays. In essence, flavopiridol ...
Today is exciting because I get to do two things. 1: Post the first successful images of West Nile virus infectined cells that I took on my own and 2: Give a basic explanation one of the most visually impressive techniques at our disposal: immunofluorescent microscopy. For those of you not familiar with fluorescent microscopy…
Endocytosis/recycling and exocytosis are mechanisms conserved through evolution allowing cells to communicate with their external medium. In order to study these dynamic processes, the present work proposes a patch-based method for detecting recycling or exocytotic events at the Plasma membrane in fast TIRF microscopy combined with the computation of normalized temporal representations of those events. Evaluation, performed on TIRF sequences showing Transferrin receptor (TfR) recycling, validates a high detection rate fully compatible with an automatic data extraction and analysis of the plasma membrane recycling process.
TY - CONF. T1 - Chirality in evanescent fields: conditions, limits and guidelines (Conference Presentation). AU - Raziman, T. V.. AU - Godiksen, Rasmus H.. AU - Müller, Moos A.. AU - Curto, Alberto G.. PY - 2020/4/1. Y1 - 2020/4/1. KW - optical chirality. KW - circular dichroism. KW - evanescent waves. KW - spin. U2 - 10.1117/12.2555726. DO - 10.1117/12.2555726. M3 - Paper. SP - 24. T2 - Metamaterials XII. Y2 - 6 April 2020 through 10 April 2020. ER - ...
Fluorescent proteins introduced or referenced by Patterson et al. (1997): Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy. Including:
We study the molecular mechanisms underlying bacterial multiplication. Bacteria are renowned for their fast multiplication. During their short cell cycles, bacteria grow, replicate and segregate their chromosomes and divide. They do all of this with sometimes blazing speed and with remarkable temporal and spatial accuracy, which explains their success. Despite the medical, agricultural and ecological importance of bacteria, little is known about the molecular mechanisms involved in growth, DNA segregation, cell division and cell morphogenesis. Similarly, the temporal and spatial mechanisms regulating these essential processes remain largely elusive. We address these fundamental questions using genetics, biochemistry, quantitative fluorescence microscopy and modeling.. ...
Dragonfly is a multi-dimensional imaging platform offering three key imaging modalities. At its core is a multi-point confocal for high-speed and high-sensitivity image capture. With speed at least 10x faster than conventional confocal technology, this mode is the optimal solution for live cell imaging, avoiding sensitivities to phototoxicity and photobleaching.. A second mode is laser-illuminated widefield epifluorescence. This mode is ideal for applications that do not benefit from confocal imaging, such as very thin samples or applications that require high laser power density, such as single molecule localisation. This mode benefits from Borealis illumination and is complemented by GPU accelerated deconvolution.. The third mode is total internal reflectance microscopy (TIRF) which is available as a factory-installed option. This is the tool of choice for imaging protein dynamics at or proximal to the cell membrane.. The Dragonfly platform is controlled by Andors Fusion software, which ...
The nature of light limits the size of the spot to which light can be focused. According to the diffraction limit a focused light distribution cannot be made smaller than approximately half of the wavelength of the used light. Uncovered in the 19th century by Ernst Abbe this has been a barrier of the achievable resolution of fluorescence light microscopes for a long time. While resolution is denoted by the ability to discern different objects of the same kind, localizing or tracking of single particles have been performed with a precision much below the diffraction limit. Several improvements in microscopy techniques have been invented in the 20th century and have resulted in increased resolution and contrast to some extent. In 1978 first theoretical ideas have been developed to break this barrier by using a 4Pi microscope as a confocal laser scanning fluorescence microscope where the light is focused ideally from all sides to a common focus which is used to scan the object by point-by-point ...
Understanding a proteins subcellular localization is critical to understanding a proteins role in the cell. The physical location of a protein limits its possible interaction partners and suggests possible biological functions for the protein [1]. The subcellular localization of a protein may be readily assessed by covalently binding it to a fluorescent protein, such as green fluorescent protein (GFP), and viewing the resulting fluorescence by microscopy. The observed pattern and intensity of fluorescence indicate the location and relative quantity of the protein in the cell.. Protein-protein interactions may be assessed via fluorescence microscopy by observing the relative subcellular localization of separate proteins simultaneously tagged with different fluorescent proteins. Proteins with strongly overlapping patterns of subcellular localization are said to colocalize; and this colocalization may indicate similar biological function or possible protein-protein interaction. Interactions may ...
Total internal reflection microscopy excites only to a depth of 100 nm, which is ideally suited to visualize fluorescent membrane proteins or proteins that operate adjacent to a membrane. Our 2 setups allow the use of UV, blue, green, yellow and red excitation, and the combination with Fluorescence Revovery after Photobleaching (FRAP). This technique uses bleaching of a small area within cells and determines protein mobility by measuring regain of fluorescence within the bleached area over time. Differential FRAP can determine association constants of protein complexes within live cells, which is done in the Flow Cytometry & Imaging Facility of the MPI Micter http://www.mpi-marburg.mpg.de/facs. Our TIRF microscopes are based on Zeiss Axioobservers, feature fast EM-CCD cameras and can also be used for regular wide field microscopy, including 3D reconstruction and time lapse microscopy.. ...
As changes in cytosolic free Ca2+ play key roles in coupling responses in neutrophils, it is important to locate and identify Ca2+ storage sites within these cells. Here, recent data is presented which highlights the functional link between microanatomical structure and cell signalling function. Fluorescent optical probes for cytosolic free Ca2+ have been used, together with organelle specific markers. We present evidence from conventional fluorescence microscopy, together with ratiometric and confocal laser scanning fluorescence microscopy, which pin-points two cellular locations for Ca2+ within the neutrophil; one within the nuclear lobes, and the other towards the cell periphery. Knowledge of these two locations provides a clear insight into how signalling in this cell type is regulated and provides a framework for explaining how specific stimuli act to produce specific responses. ...
Skeletal muscle. Rotating fluorescence light microscopy footage of skeletal muscle tissue from a tongue. Fluorescent dyes have been used to highlight tissues, cellular structures and proteins. Proteins highlighted include actin (green). - Stock Video Clip K004/6259
Blood vessel. Rotating fluorescence light microscopy footage of a blood vessel (pink) traversing skeletal muscle in tissue from the tongue. Fluorescent dyes have been used to highlight tissues, cellular structures and proteins. - Stock Video Clip K004/6256
Fluorescence microscopy applications are rapidly expanding in many medical and biological research laboratories. The mainstream of fluorescence microscopy has undergone an almost total shift from utilizing transmitted light to incident light, accompanied by the introduction of many new and different fluorochromes. Fluorescence microscopy applications are rapidly expanding in many medical and biological research laboratories. The mainstream of fluorescence microscopy has undergone an almost total shift from utilizing transmitted light to incident light, accompanied by the introduction of many new and different fluorochromes.
Fluorescence is one of the most commonly used physical phenomena in biological and analytical microscopy, mainly because of its high sensitivity and high specificity. Fluorescence is a form of luminescence. Fluorescence microscopy even allows users to determine the distribution of a single molecule species, its amount and its localization inside a cell. Colocalization and interaction studies can be performed, and ion concentrations, as well as intra- and intercellular processes like endocytosis and exocytosis, can be observed. With the help of super-resolution fluorescence microscopy, it is even possible to image sub-resolution structures.. ...
Bayesian localization microscopy reveals nanoscale podosome dynamics. Susan Cox, Edward Rosten, James Monypenny, Tijana Jovanovic-Talisman, Dylan T Burnette, Jennifer Lippincott-Schwartz, Gareth E Jones & Rainer Heintzmann. Nature Methods. 2012, volume 9: 195-200. doi: 10.1038/nmeth.1812. http://f1000.com/13978956 An analytical approach based on the concept of fluorophore localisation provides dynamic super-resolution data of xFP- labelled live cells using a common arc lamp based wide-field fluorescence microscope. One method of achieving fluorescence super-resolution is based around finding the positions of fluorescent molecules that label the cellular structure of interest. In this approach, positions can be determined precisely and accurately using fluorescent probes that can be photoactivated, photoconverted or photoswitched to generate single images with emitter densities of only about one active fluorophore per diffraction-limited area. Many images each containing subsets of active ...
Bayesian localization microscopy reveals nanoscale podosome dynamics. Susan Cox, Edward Rosten, James Monypenny, Tijana Jovanovic-Talisman, Dylan T Burnette, Jennifer Lippincott-Schwartz, Gareth E Jones & Rainer Heintzmann. Nature Methods. 2012, volume 9: 195-200. doi: 10.1038/nmeth.1812. http://f1000.com/13978956 An analytical approach based on the concept of fluorophore localisation provides dynamic super-resolution data of xFP- labelled live cells using a common arc lamp based wide-field fluorescence microscope. One method of achieving fluorescence super-resolution is based around finding the positions of fluorescent molecules that label the cellular structure of interest. In this approach, positions can be determined precisely and accurately using fluorescent probes that can be photoactivated, photoconverted or photoswitched to generate single images with emitter densities of only about one active fluorophore per diffraction-limited area. Many images each containing subsets of active ...
Define wide-field fluorescence microscopy: Wide-field fluorescence microscopy uses either naturally occurring structures or staining with fluorescent ...
Light Microscopy and Resolution -- Super-Resolution Microscopy: Principles, Techniques, and Applications -- Foundations of STED Microscopy -- Application of Real-Time STED Imaging to Synaptic Vesicle Motion -- Photoactivated Localization Microscopy for Cellular Imaging -- Data Analysis for Single-Molecule Localization Microscopy -- Super-Resolution Fluorescence Microscopy Using Structured Illumination -- Application of Three-Dimensional Structured Illumination Microscopy in Cell Biology: Pitfalls and Practical Considerations -- Scanning Near-Field Optical Microscopy for Investigations of Bio-Matter -- Atomic Force Microscopy of Living Cells -- X-Ray Microscopy for Neuroscience: Novel Opportunities by Coherent Optics -- Nonlinear Optics Approaches Towards Sub-Diffraction Resolution in CARS Imaging -- Photo-Oxidation Microscopy: Bridging the Gap Between Fluorescence and Electron Microscopy -- Requirements for Samples in Super-Resolution Fluorescence Microscopy -- Probing Biological Samples in ...
Fingerprint Dive into the research topics of A fluorescence microscopy study of quantum dots as fluorescent probes for brain tumor diagnosis. Together they form a unique fingerprint. ...
This dissertation describes the application of fluorescence microscopy techniques to investigations of mass transport phenomena in self-assembled nanomaterials. The microscopic morphologies of the materials and the mass-transport dynamics of probe molecules dispersed within them were assessed with high temporal and spatial resolution by single molecule imaging and spectroscopic methods. Three distinct sets of experiments were performed in completing the work for this dissertation. In the first study, single molecule imaging was employed to explore the interactions and field-induced migration of double-stranded DNA (ds-DNA) molecules with nanostructured Pluronic F127 gels. While DNA interactions with nanostructured gels have been explored in the past, none had apparently looked at these interactions in gels comprising hexagonally ordered arrays of cylindrical micelles. Therefore, these studies focused on materials DNA dispersed in flow aligned hexagonal F127. DNA molecules were found to be ...
Super-resolution microscopy has tremendously progressed our understanding of cellular biophysics and biochemistry. Specifically, 4pi fluorescence microscopy technique stands out because of its axial super-resolution capability. All types of 4pi-microscopy techniques work well in conjugation with deconvolution techniques to get rid of artifacts due to side-lobes. In this regard, we propose a technique based on spatial filter in a 4pi-type-C confocal setup to get rid of these artifacts. Using a special spatial filter, we have reduced the depth-of-focus. Interference of two similar depth-of-focus beams in a 4 pi geometry result in substantial reduction of side-lobes. Studies show a reduction of side-lobes by 46% and 76% for single and two photon variant compared to 4pi - type - C confocal system. This is incredible considering the resolving capability of the existing 4pi - type - C confocal microscopy. Moreover, the main lobe is found to be 150 nm for the proposed spatial filtering technique as ...
Total Internal Reflection Fluorescence Microscopy (TIRFM) is a common method to study structural and dynamic processes at the single-molecule level. The principle is based on an optical phenomenon where electromagnetic radiation is not refracted but totally reflected on the boundary surface of two different media. An evanescent field is generated at the boundary surface and penetrates the medium with lower optical density where fluorescent molecules can be excited. This evanescent wave attains its maximum intensity at the surface and then exponentially decays with increasing penetration depth. Therefore, only molecules located near the surface are excited, as opposed to molecules situated at distances exceeding approx. 100 nm, and thus TIRF efficiently decreases background signals of the bulk solution and allows sensitive detection down to single molecules ...
Part of the prestigious Max Planck Society based in Germany, MPFI is the first and only institute of its kind in North America. Situated in the new biosciences cluster in scenic Palm Beach County in South Florida, MPFI provides a vibrant, collaborative environment where scientists are provided generous ongoing support to conduct high impact research at the cutting edge.. ...
Box 1. Determining the quality of a TIRF set up. The test samples described below can be used to check the quality of any TIRF set up. They should always be prepared using a cover slip with the correct thickness and refractive index for the objective.. Test samples. Fluorescent microbeads. These can be purchased from many sources, including Invitrogen (Carlsbad, CA) and Bangs Laboratories (Fishers, IN). The beads should be of subresolution size (100 nm diameter or less), and selected to have excitation and emission spectra that match typical experimental conditions. The beads should be diluted in water and applied to the cover slip. PBS can be added to increase the number of beads that adhere to the surface.. DiI. A convenient, uniform, fluorescent film can be easily made on a cover slip surface with the lipophilic fluorophore DiI (Invitrogen, Carlsbad, CA). Dissolve the DiI at 0.5 mg/ml in ethanol and place a single droplet of the solution on a glass cover slip. Then, before the solution dries, ...
Super-resolution fluorescence imaging methods based on reversible photoswitching of fluorophores with subsequent localization currently develop to promising tools for cellular imaging. Since most of these methods rely on the transfer of the majority of fluorophores to a non-fluorescent dark state and precise localization of separated fluorescent fluorophores, the photophysical properties of photoswitchable fluorophores have to be carefully controlled. The achievable resolution and herewith the ability to resolve a structural feature depends not only on the brightness of the fluorophores, but also on the labeling density and on the stability or lifetime of the non-fluorescent dark state. Here, we discuss how the ratio of off- and on-switching of a fluorophore affects resolution. We compare experimental data with theoretical simulations and present a strategy to customize photoswitching characteristics to achieve optimal optical resolution. (C) 2010 Elsevier B.V. All rights reserved ...
The ASU Library acknowledges the twenty-two Native Nations that have inhabited this land for centuries. Arizona State Universitys four campuses are located in the Salt River Valley on ancestral territories of Indigenous peoples, including the Akimel Oodham (Pima) and Pee Posh (Maricopa) Indian Communities, whose care and keeping of these lands allows us to be here today. ASU Library acknowledges the sovereignty of these nations and seeks to foster an environment of success and possibility for Native American students and patrons. We are advocates for the incorporation of Indigenous knowledge systems and research methodologies within contemporary library practice. ASU Library welcomes members of the Akimel Oodham and Pee Posh, and all Native nations to the Library.. ...
A super-resolution platform for correlative live single-molecule imaging and STED microscopy. Super-resolution microscopy offers tremendous opportunities to unravel the complex and dynamic architecture of living cells. However, current super-resolution microscopes are well suited for revealing protein distributions or cell morphology, but not both. We present a super-resolution platform that permits correlative single-molecule imaging and stimulated emission depletion microscopy in live cells. It gives nanoscale access to the positions and movements of synaptic proteins within the morphological context of growth cones and dendritic spines.. ...
0050] This species of surgical fluorescence stereomicroscopes requires specific properties known to one skilled in the art, and specific constituents of the microscope that make said properties possible. Those pertinent to conventional fluorescence microscopy (which already existed long before the creation of surgical fluorescence stereomicroscopes) are: [0051] an excitation light source or excitation illumination device (formerly, for example, often a mercury vapor lamp), [0052] an excitation filter to improve the quality of the excitation light (spectrally filtering out those light wavelength regions that do not contribute particularly well, or at all, to fluorescence excitation), and a blocking filter or observation filter in the observation beam path of the surgical fluorescence stereomicroscope. The latter serves in turn to filter out the excitation light to a greater or lesser extent, since little or none of it is after all intended in principle to be seen, but the emission of the ...
When Eric Betzig and colleagues first described their new microscopy method, PALM, they chose to highlight its power by comparing it to an ultra-high-resolution approach: transmission electron microscopy (TEM). PALM, or photoactivated localization microscopy, is a super-resolution fluorescence technique allowing users to circumvent the 200 nm diffraction limit that constrains optical microscopy, mapping fluorophores to within…
Day 1. Cell growth. MCF7, LLC-PK1 cells. Grow MCF7 or LLC-PK1 cells at 37˚C in high glucose DMEM, supplemented with 8% FBS, sodium pyruvate and antibiotics [15, 16]. 24 hours prior to the experiment, trypsinize cells and culture overnight on glass cover slips coated with poly-L-lysine.. Superior cervical ganglion (SCG) neurons. Prepare SCG neurons from postnatal day 1 to 7 as described [11]. Culture cells on glass coverslips (coated with laminin) in modified 35 mm dishes (Corning) at 37˚C for 7-10 days; change medium twice per week.. Day 2. Treatment with MitoTracker®. Remove medium and treat cells with fresh DMEM supplemented with MitoTracker® (diluted 1:5,000 in DMEM). Incubate for 30 minutes at 37˚C in a cell culture incubator (5% CO2).. Fixation, permeabilization, block of non-specific binding sites * MCF7 and LLC-PK1 cells*. After 30 min incubation with MitoTracker®, remove the tissue culture medium, wash cells once with pre-warmed PBS. Fix samples in 3.7 % formaldehyde/PBS for 15 min ...
Structural characterization by super-resolution microscopy has become increasingly widespread, particularly in the biological community. The technique is powerful because it can produce real-space images with resolutions of tens of nanometers, while sample preparation is relatively non-invasive. Previous studies have applied these techniques to important scientific problems in the life sciences, but relatively little work has explored the attainable limit of resolution using samples of known structure. In this work, we apply photo-activated localization microscopy (PALM) to polymer films that have been nanopatterned using electron-beam lithography. Trace amounts of a rhodamine spiroamide dye are dispersed into nanostructured poly(methyl methacrylate), and UV-induced switching of the fluorophores enables nanoscale localization of single molecules to generate a final composite super-resolution image. Features as small as 25 nm half-pitch are clearly resolvable.. We also explore the effect of ...
Fluorescence filters help to isolate specific wavelengths of fluoresced light - which might seem light years away from anything to do with jellyfish evolution.. But in fact, many modern scientific processes are derived from naturally occurring biochemical capabilities, such as the green fluorescent proteins (GFPs) found off the coast of North America in the jellyfish species Aequorea victoria.. These green proteins can be used as fluorescent markers for biological phenomena, by attaching them as genetic tags and then using fluorescence filters to see where they go.. Some GFPs can do more though - including changing colour to become red - and these are especially valuable for super-resolution fluorescence microscopy.. A team at Arizona State University have now discovered a hinge migration mechanism that they say is the key for evolution of a green-to-red photoconvertible phenotype in a GFP.. It took eight years to unlock this process, which is described in a paper published in the academic ...
Visualizing 3D structure and dynamics at the molecular scale is a current and critical need in biomedical research. Many sub-cellular features, for example the morphology of many organelles or the 3D organization of chromatin, cannot be resolved by standard light microscopy. Improving the resolution of light microscopy has therefore been an urgent need of biological research for many decades. Today, several methods achieve sub-100 nm resolution by taking advantage of reversible or irreversible photo-physical switching properties of fluorescent markers. Our research group in the Department of Cell Biology at Yale University School of Medicine is developing new fluorescence microscopy techniques with spatial and/or temporal resolutions exceeding far beyond current technology and also applying them to a diverse set of biological questions.. Specialized Terms: Super-resolution fluorescence ...
-A key step of adaptive immune responses is the T lymphocyte capacity to detect the presence of foreign antigens on specialized cells with high speed and specificity during contacts lasting a few minutes. Much evidence suggests that there is a deep link between the lifetime of molecular interactions between T cell receptors and ligands and T cell activation, but the precise mechanisms of bond formation and dissociation remain incompletely understood. Previous experiments done with interference reflection microscopy/reflection interference contrast microscopy disclosed transverse motions with several nanometer average amplitude of micrometer size membrane zones. More recently, total internal reflection fluorescence microscopy was used to show that the initial interaction between primary T lymphocytes and model surfaces involved the tip of microvilli (typically 0.2 lm 2 area) generating apparent contacts of a few seconds that allowed cells to detect ligands of their membrane receptors. Here we show that
The study of protein dynamics is essential for understanding the multi-molecular complexes at subcellular levels. Fluorescent Protein (XFP)-tagging and time-lapse fluorescence microscopy enable to observe molecular dynamics and interactions in live cells, unraveling the live states of the matter. Original image analysis methods are then required to process challenging 2D or 3D image sequences. Recently, tracking methods that estimate the whole trajectories of moving objects have been successfully developed. In this paper, we address rather the detection of meaningful events in spatio-temporal fluorescence image sequences, such as apparent stable stocking areas involved in membrane transport. We propose an original patch-based Markov modeling to detect spatial irregularities in fluorescence images with low false alarm rates. This approach has been developed for real image sequences of cells expressing XFP-tagged Rab proteins, known to regulate membrane trafficking.
Our other large Zeiss epifluorescent microscope is the Zeiss Axiophot. In addition to fluorescence microscopy, this microscope is also used for phase contrast and differential interference contrast (DIC) microscopy. Though it is an older scope, it was recently outfitted with a modern digital Zeiss AxioCam MR camera and utilizes the same Zen software as the new Axio Imager 2. Phase contrast and DIC microscopy allow for observation of unstained, live cells and structures. For fluorescent microscopy, this microscope utilizes a Zeiss XBO 75 (75W xenon arc lamp) and HBO 100 (100W mercury arc lamp).. For additional information including manuals, manufacturer links, protocols and publications, see our resources page. To see images captured by this microscope, see the gallery.. ...
Find Molecular Probes® fluorescent labels for multiplexed super-resolution microscopy (SRM) applications, including STORM, STED, SIM, and two-photon microscopy.
phdthesis{f9c27a48-2eb3-4555-b7c6-9d9c5fb019ce, abstract = {In the research area of cleaning and process hygiene it is important to understand the underlying mechanisms behind the deposition of particles and macromolecules onto surfaces. The deposition process is described as the transport of particles to the surface followed by attachment. If the hydrodynamic forces are strong enough, the particles can be re-entrained.,br/,,br, ,br/,,br, In this work particle deposition was studied <i>in situ</i>. In order to obtain a well-defined mass transfer to a glass surface covered with indium-tin oxide onto which deposition was to occur, an experimental setup consisting of a wall-jet cell was employed. Model solutions consisting of polystyrene latex particles of two particle radii, 0.23 um and 0.38 um, were used in the deposition experiments. The deposited particles were visualized by total internal reflection microscopy. The construction of the cell made it possible to radially scan the ...
2. What did the paper show that normal microscopy could not show. In this paper Schermelleh et al. noted that although normal fluorescence light microscopy permits the cellular components to be visualised in various colours, it is limited in providing adequate resolution. They overcame this problem by applying the use of 3D-SIM. In this paper they use it to study the mammalian nucleus. In particular, with the use of 3D-SIM they were able to resolve single NPCs[1], differentially localize distinct NPC components[2] and detect double-layered invaginations of the nuclear envelope[3], all of which would not be possible with the use of conventional microscopy. Schermelleh et al. notes that the use 3D-SIM will allow for a greater exploration of the subcellular structures beyond the diffraction limit of the emitted light[4] Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy. --Z3254598 00:52, 22 March 2012 (EST) ...
This course will cover the theory and practical application of current super-resolution microscopy techniques to biological questions.
We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer superior structural preservation, performing in-resin super-resolution, however, remains a challenge. We identified key aspects of the sample preparation procedure of high pressure freezing, freeze substitution and resin embedding that are critical for preserving fluorescence and photo-switching of standard fluorescent proteins, such as mGFP, mVenus and mRuby2. This enabled us to combine single molecule localization microscopy with transmission electron microscopy imaging of standard fluorescent proteins in cryo-fixed resin embedded cells. We achieved a structural resolution of 40-50 nm (~17 nm average single molecule localization accuracy) in the fluorescence images without the use of chemical fixation or special fluorophores. Using this approach enabled the correlation of fluorescently labeled ...
The cytoskeleton is involved in many cellular processes. Over the last decade, super-resolution microscopy has become widely available to image cytoskeletal structures, such as microtubules and actin, with great detail. For example, Single-Molecule Localization Microscopy (SMLM) achieves resolutions of 5-50 nm through repetitive sparse labeling of samples, followed by Point-Spread-Function ... read more analysis of individual fluorophores. Whereas initially this approach depended on the controlled photoswitching of fluorophores targeted to the structure of interest, alternative techniques now depend on the transient binding of fluorescently labeled probes, such as the small polypeptide lifeAct that can transiently interact with polymerized actin. These techniques allow for simple multicolor imaging and are no longer limited by a fluorophores blinking properties. Here we describe a detailed step-by-step protocol to purify, label, and utilize the lifeAct fragment for SMLM. This purification and ...
RhoGTPases are important regulators of the cell cytoskeleton, controlling cell shape, migration and proliferation. Previously we showed that ARHGAP18 in endothelial cells is important in cell junctions. Here we show, using structured illumination microscopy (SIM), ground-state depletion (GSD), and total internal reflection fluorescence microscopy (TIRF) that a proportion of ARHGAP18 localizes to microtubules in endothelial cells, as well as in nonendothelial cells, an association confirmed biochemically. In endothelial cells, some ARHGAP18 puncta also colocalized to Weibel-Palade bodies on the microtubules. Depletion of ARHGAP18 by small interfering RNA or analysis of endothelial cells isolated from ARHGAP18-knockout mice showed microtubule destabilization, as evidenced by altered morphology and decreased acetylated α-tubulin and glu-tubulin. The destabilization was rescued by inhibition of ROCK and histone deacetylase 6 but not by a GAP-mutant form of ARHGAP18. Depletion of ARHGAP18 resulted ...
Project Description: The Kner and De La Fuente Labs are interested in using a novel 3D superresolution fluorescence microscopy approach to study nucleosome organization and the role of specific proteins in chromatin organization. The project involves the engineering of a new superresolution microscope, imaging of chromosomes, and biological questions that are important for in reproduction.. REU Student Role and Responsibility: The REU student will assist a graduate student in developing the microscope and in imaging experiments on samples form the De La Fuente lab. The student will assist in developing software to run the microscope and process the images, will help run the microscope to collect data, and will help prepare the samples for imaging.. Expected Outcome for REU student: The student will be trained in cutting-edge microscopy techniques and exposed to important questions in modern biology. Superresolution imaging of the nucleus is a hot topic. This project is expected to result in at ...
Endocrinology. 2003 Aug;144(8):3532-40. Ovine placental lactogen-induced heterodimerization of ovine growth hormone and prolactin receptors in living cells is demonstrated by fluorescence resonance energy transfer microscopy and leads to prolonged phosphorylation of signal transducer and activator of transcription (STAT)1 and STAT3. Biener E, Martin C, Daniel N, Frank SJ, Centonze VE, Herman B, Djiane J, Gertler A ...
Researchers from UCL, the National Physical Laboratory and the Royal Free Hospital have differentiated between patients with a rare bleeding disorder and healthy volunteers using super-resolution microscopy, providing an alternative method for accurately and cost-effectively diagnosing rare platelet diseases.
Get an introduction to the technologies, unique benefits, practical examples, and future developments of super-resolution microscopy.
Repulsive guidance molecules (RGMs) control crucial processes including cell motility, adhesion, immune-cell regulation and systemic iron metabolism. RGMs signal via the neogenin (NEO1) and the bone morphogenetic protein (BMP) pathways. Here, we report crystal structures of the N-terminal domains of all human RGM family members in complex with the BMP ligand BMP2, revealing a new protein fold and a conserved BMP-binding mode. Our structural and functional data suggest a pH-linked mechanism for RGM-activated BMP signaling and offer a rationale for RGM mutations causing juvenile hemochromatosis. We also determined the crystal structure of the ternary BMP2-RGM-NEO1 complex, which, along with solution scattering and live-cell super-resolution fluorescence microscopy, indicates BMP-induced clustering of the RGM-NEO1 complex. Our results show how RGM acts as the central hub that links BMP and NEO1 and physically connects these fundamental signaling pathways. Repulsive guidance molecule is a structural ...
Extraordinary transmission based axial imaging (EOT-AIM) for cell microscopy is reported. EOT-AIM uses linear arrays of nanoapertures, each of which samples target fluorescence up to a preset axial distance from surface, in combination with wide-field microscopy for acquisition of lateral images. Current design of nanoapertures provides EOT-AIM with axial super-resolution that is as small as 20 nm for a depth range of 500 nm. Experiments were performed for the measurement of the axial distribution of ganglioside in mouse macrophage (RAW264.7) cells using FITC-conjugated cholera toxin subunit B. The results were successfully confirmed with conventional confocal and total internal reflection fluorescence microscopy. ...
Definition of Evanescent wave in the Legal Dictionary - by Free online English dictionary and encyclopedia. What is Evanescent wave? Meaning of Evanescent wave as a legal term. What does Evanescent wave mean in law?
Recently implemented fluorescence imaging techniques, such as total internal reflection fluorescence microscopy and two-photon laser scanning micro-scopy, have made possible multiscale analysis of the immune response from single molecules in an interface to cells moving in lymphoid tissues and tumors. In this review, we consider components of T cell sensitivity: the immunological synapse, the coordination of migration, and antigen recognition in vivo. Potency, dose, and detection threshold for peptide-MHC determine T cell sensitivity. The immunological synapse incorporates T cell receptor microclusters that initiate and sustain signaling, and it also determines the positional stability of the T cells through symmetry and symmetry breaking. In vivo decisions by T cells on stopping or migration are based of antigen stop signals and environmental go signals that can sometimes prevent arrest of T cells altogether, and thus can change the outcome of antigen encounters. © 2010 Springer-Verlag Berlin
Recently implemented fluorescence imaging techniques, such as total internal reflection fluorescence microscopy and two-photon laser scanning micro-scopy, have made possible multiscale analysis of the immune response from single molecules in an interface to cells moving in lymphoid tissues and tumors. In this review, we consider components of T cell sensitivity: the immunological synapse, the coordination of migration, and antigen recognition in vivo. Potency, dose, and detection threshold for peptide-MHC determine T cell sensitivity. The immunological synapse incorporates T cell receptor microclusters that initiate and sustain signaling, and it also determines the positional stability of the T cells through symmetry and symmetry breaking. In vivo decisions by T cells on stopping or migration are based of antigen stop signals and environmental go signals that can sometimes prevent arrest of T cells altogether, and thus can change the outcome of antigen encounters. © 2010 Springer-Verlag Berlin
BILLERICA -- Bruker Corp. recently announced that it has acquired Vutara Inc., a Salt Lake City-based provider of super-resolution fluorescence microscopy for life-science applications.
Scattering-Assisted Localization Microscopy (SALM) is a super-resolution microscopy tool where high resolution is obtained based on the size of the grain in the speckle pattern on the sample.
Researchers measured times from cytokinesis to budding (G1) and from budding to cytokinesis in haploids, diploids or tetraploids (mothers and daughters), using time-lapse fluorescence microscopy of strains expressing Myo1 tagged with green fluorescent protein (Myo1-GFP). Standard methods were used throughout strain and plasmid constructions. All strains are W303-congenic. All integrated constructs were characterized by Southern blot analysis. Cells were prepared for time-lapse microscopy as described (Bean et al. 2006 PMID 16387649). Researchers observed growth of microcolonies with fluorescence time-lapse microscopy at 30?°C using a Leica DMIRE2 inverted microscope with a Ludl motorized XY stage. Images were acquired every 3?min for cells grown in glucose and every 6?min for cells grown in glycerol/ethanol with a Hamamatsu Orca-ER camera. They used custom Visual Basic software integrated with ImagePro Plus to automate image acquisition and microscope control ...
Impaired insulin secretion in type 2 diabetes (T2D) is linked to reduced insulin granule docking, disorganization of the exocytotic site, and impaired glucose-dependent facilitation of insulin exocytosis. We show in β cells from 80 human donors that the glucose-dependent amplification of exocytosis is disrupted in T2D. Spatial analyses of granule fusion events, visualized by total internal reflection fluorescence microscopy in 24 of these donors, demonstrated that these events are nonrandom across the surface of β cells from donors with no diabetes. The compartmentalization of events occurs within regions defined by concurrent or recent membrane-resident secretory granules. This organization, and the number of membrane-associated granules, is glucose dependent and notably impaired in T2D β cells. Mechanistically, multichannel Kv2.1 clusters contribute to maintaining the density of membrane-resident granules and the number of fusion hotspots, while SUMOylation sites at the channel N- (K145) ...
Impaired insulin secretion in type 2 diabetes (T2D) is linked to reduced insulin granule docking, disorganization of the exocytotic site, and impaired glucose-dependent facilitation of insulin exocytosis. We show in β cells from 80 human donors that the glucose-dependent amplification of exocytosis is disrupted in T2D. Spatial analyses of granule fusion events, visualized by total internal reflection fluorescence microscopy in 24 of these donors, demonstrated that these events are nonrandom across the surface of β cells from donors with no diabetes. The compartmentalization of events occurs within regions defined by concurrent or recent membrane-resident secretory granules. This organization, and the number of membrane-associated granules, is glucose dependent and notably impaired in T2D β cells. Mechanistically, multichannel Kv2.1 clusters contribute to maintaining the density of membrane-resident granules and the number of fusion hotspots, while SUMOylation sites at the channel N- (K145) ...
Fluorescence Microscope for sale, new Infinitive Plan Phase contrast Microscope Inverted Fluorescence Microscope CE A16.1023 of Opto-Edu (Beijing) Co., Ltd. from China.
We are a little late to the party (it has been a busy month at Protocols HQ!), but Nature Protocols would like to extend their warmest congratulations to all this years winners of the Nobel Prize for Chemistry: Eric Betzig, Stefan Hell and William E. Moerner for their contributions to the development of super-resolution microscopy.. We are very pleased to have published a Nature Protocol from the Betzig lab earlier this year on Bessel beam plane illumination microscopy. And we are equally pleased that the Protocol Exchange can claim a Nobel Laureate amongst its authors; the Moerner lab has published a guide to using Easy-DHPSF to measure the precise localisations of molecules in images acquired using a wide-field DH epifluorescence microscope. I would also encourage you to visit Moerners very informative lab website, if only to find out about the guacamole!. ...
If you are looking for a Distributor of Olympus IX83 Microscope in India then you need to look no further than DSS Image. The fully motorized and automated inverted microscope system IX83 is the most advanced in the IX3 series of inverted imaging systems
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By surpassing Abbes diffraction limit, super-resolution microscopy (SRM) has opened the door to the visualization and the understanding of complex structures in the 1-100 nm range. SRM combines typical features of optical microscopy (low invasiveness, high penetration depth, high sensitivity, chemical specificity, straightforward sample preparation) with nanometer spatial resolution. Despite its widespread in cell-biology and bio-physics, SRM is still in its infancy for in-depth visualization of mesoscale features in man-made materials. At the VoetsLab, we use SRM techniques such as PAINT (point accumulation for imaging in nanoscale topography), PALM (photo-activatable localization microscopy) and STORM (stochastic optical reconstruction microscopy) to visualize in-situ structural composition, stability, exchange dynamics and growth direction of self-assembled architectures ranging from one-dimensional fibers to multi-dimensional colloids. By doing so, we aim for a more rational and ...
The assembly process of the human immunodeficiency virus 1 (HIV-1) is driven by the viral polyprotein Gag. Fluorescence imaging of Gag protein fusions is widely performed and has revealed important information on viral assembly. Gag fusion proteins are commonly co-transfected with an unlabeled form of Gag to prevent labeling artifacts such as morphological defects and decreased infectivity. Although viral assembly is widely studied on individual cells, the efficiency of the co-transfection rescue has never been tested at the single cell level. Here, we first develop a methodology to quantify levels of unlabeled to labeled Gag in single cells using a fluorescent reporter protein for unlabeled Gag and fluorescence correlation spectroscopy. Using super-resolution imaging based on photoactivated localization microscopy (PALM) combined with molecular counting we then study the nanoscale morphology of Gag clusters as a function of unlabeled to labeled Gag ratios in single cells. We show that for a ...
Yuste R (Dec 2005). "Fluorescence microscopy today". Nature Methods. 2 (12): 902-4. doi:10.1038/nmeth1205-902. PMID 16299474. ... The availability of GFP and its derivatives has thoroughly redefined fluorescence microscopy and the way it is used in cell ... This has triggered the development of highly automated live-cell fluorescence microscopy systems, which can be used to observe ... Zn(II) binding increases fluorescence intensity, while Cu(II) binding quenches fluorescence and shifts the absorbance maximum ...
Yuste R (2005). "Fluorescence microscopy today". Nature Methods. 2 (12): 902-904. PMID 16299474. doi:10.1038/nmeth1205-902.. ... Cryoelectron microscopy is used to produce lower-resolution structural information about very large protein complexes, ... Finally, the gold-standard method of cellular localization is immunoelectron microscopy. This technique also uses an antibody ... For example, indirect immunofluorescence will allow for fluorescence colocalization and demonstration of location. Fluorescent ...
Fluorescence Microscopy. Cambridge University Press. p. 22. ISBN 0-521-23641-X. Archived from the original on November 13, 2012 ... Practical Fluorescence, Second Edition, Marcel Dekker, Inc., New York. ISBN 0-8247-8350-6. Banwell C.N. and McCash E.M. ... This number varies over several orders of magnitude, depending on the sample, and is known as the fluorescence lifetime of the ... Principles of Fluorescence Spectroscopy, Plenum Press, New York. ISBN 0-387-31278-1. Guilbault, G.G. 1990. ...
In fluorescence microscopy objects out of the focal plane only interfere with the image if they are illuminated and fluoresce. ... Lightsheet based fluorescence microscopy illuminates the sample with excitation light under an angle of 90° to the direction of ... Gratton E, Barry NP, Beretta S, Celli A (September 2001). "Multiphoton fluorescence microscopy". Methods. 25 (1): 103-10. doi: ... Total internal reflection microscopy is a fluorescent microscopy technique, which intentionally restricts observation to either ...
Yuste R (December 2005). "Fluorescence microscopy today". Nature Methods. 2 (12): 902-4. doi:10.1038/nmeth1205-902. PMID ... Cryoelectron microscopy is used to produce lower-resolution structural information about very large protein complexes, ... This technique also uses an antibody to the protein of interest, along with classical electron microscopy techniques. The ... In more recent times, cryo-electron microscopy of large macromolecular assemblies and computational protein structure ...
Rusk, N. (2009). "The fluorescence microscope". Milestones in Light Microscopy. Nature Publishing Group. Heimstädt O. (1911). " ... Fortunately, Köhler saw the potential of fluorescence. A filtering technique for fluorescence excitation light was developed by ... In 1973 Steinkamp and the team at Los Alamos follow up with a fluorescence-based cell sorter. In 1978, at the Conference of the ... In the late 1960s, Van Dilla at Los Alamos National Laboratory built the first non microscopy-based cytophotometer. He did this ...
... microscopy, saturated structured-illumination microscopy (SSIM), fluorescence photoactivation localization microscopy (FPALM), ... extraneous undesired specific fluorescence, and nonspecific fluorescence. Autofluorescence includes fluorescence emitted from ... Immunofluorescence is a technique used for light microscopy with a fluorescence microscope and is used primarily on ... Huang, Bo; Bates, Mark; Zhuang, Xiaowei (2009-06-02). "Super-Resolution Fluorescence Microscopy". Annual Review of Biochemistry ...
... can be visualised with fluorescence microscopy by using aequorin as a reporter protein. The ... Axelrod, D. (2008). Total Internal Reflection Fluorescence Microscopy. In J. J. Correia & H. W. Detrich (Eds.), Biophysical ... Dense core granule exocytosis viewed by total internal reflection fluorescence microscopy". Methods. 46 (3): 233-238. doi: ... microdomains at the uropod of living morphologically polarized human neutrophils using flash lamp-based fluorescence microscopy ...
To minimize damage to the visualized cells, two-photon microscopy is often invoked to detect the fluorescence from the ... "Optimizing 3D multiphoton fluorescence microscopy". Optics Letters. 38 (19): 3945-3948 Miyawaki A, Llopis J, Heim R, McCaffery ... which is simply the ratio of the change in fluorescence (F - F0) over the baseline fluorescence. This can be related to the SNR ... Light field microscopy methods extend functional readout of neural activity capabilities in 3D volumes. Cannell MB, Berlin JR, ...
For quantum dots, prolonged single-molecule microscopy showed that 20-90% of all particles never emit fluorescence. On the ... Fluorescent dyes Fluorescence in the life sciences Quenching of fluorescence Fluorescence recovery after photobleaching (FRAP ... Juan Carlos Stockert, Alfonso Blázquez-Castro (2017). "Chapter 3 Dyes and Fluorochromes". Fluorescence Microscopy in Life ... Fluorophores can also be used to quench the fluorescence of other fluorescent dyes (see article Quenching (fluorescence)) or to ...
In fluorescence microscopy, colocalization refers to observation of the spatial overlap between two (or more) different ... Colocalization is used in real-time single-molecule fluorescence microscopy to detect interactions between fluorescently ... Curr Protoc Cell Biol "Quantitative colocalization analysis of confocal fluorescence microscopy images." Archived 2009-11-28 at ... 2013). "Bridging the gap between qualitative and quantitative colocalization results in fluorescence microscopy studies". Sci ...
A QD-LED integrated at a scanning microscopy tip was used to demonstrate fluorescence near-field scanning optical microscopy ( ... Juan Carlos Stockert, Alfonso Blázquez-Castro (2017). "Chapter 18 Luminescent Solid-State Markers". Fluorescence Microscopy in ... This is called fluorescence. In a simplified model, the energy of the emitted photon can be understood as the sum of the band ... To improve fluorescence quantum yield, quantum dots can be made with shells of a larger bandgap semiconductor material around ...
Fluorescence Microscopy in Life Sciences. Bentham Science Publishers. ISBN 978-1-68108-519-7. Retrieved 17 December 2017. CS1 ...
Analysis by flow cytometry and fluorescence microscopy". Journal of Immunological Methods. 133 (1): 87-97. doi:10.1016/0022- ... due to the progressive halving of CFSE fluorescence within daughter cells following each cell division.[3] The only limitation ... approximately 7-8 cell divisions can be identified before the CFSE fluorescence is too low to be distinguished above the ...
CS1 maint: discouraged parameter (link) Hepler, P. K.; B. E. S. Gunning (1998). "Confocal fluorescence microscopy of plant ... Building on the work of Shinya Inoué and Andrew Bajer using polarized light microscopy, Hepler used electron microscopy to ... Zeiger, E.; P. K. Hepler (1979). "Blue light-induced, intrinsic vacuolar fluorescence in onion guard cells". Journal of Cell ... S2CID 19236616.CS1 maint: multiple names: authors list (link) "Poems and Quotations About the MicroWorld". Microscopy Society ...
Denk, W.; Strickler, J.; Webb, W. (1990). "Two-photon laser scanning fluorescence microscopy". Science. 248 (4951): 73-76. ... Another technique, the Two Photons Microscopy (2P), invented by Winfried Denk (for which he has been awarded the Brain Prize in ...
"Molecular Expressions Microscopy Primer: Specialized Microscopy Techniques - Fluorescence Digital Image Gallery - Male Rat ... "Olympus Microscopy Resource Center , Fluorescence Digital Image Gallery - Epithelial Cells". Retrieved 17 December 2013. ... Brescia, Jr, Peter J.; Banks, Peter (24 June 2014). "High Resolution Fluorescence Microscopy of PtK2 Cells Undergoing Mitosis ...
2012). "Circular Dichroism Probed by Two-Photon Fluorescence Microscopy in Enantiopure Chiral Polyfluorene Thin Films". J. Am. ... Denk, W.; Strickler, J.; Webb, W. (1990). "Two-Photon Laser Scanning Fluorescence Microscopy". Science. 248 (4951): 73-76. ... The first experimental measurement of TPCD was performed in 1995 using a fluorescence based technique (FD-TPCD), but it was not ... Gunde, K.E.; Richardson, F.S. (1995). "Fluorescence-Detected Two-Photon Circular Dichroism of Gd3+ in Trigonal Na3[Gd(C4H4O5)3 ...
Two-photon laser scanning fluorescence microscopy Denk 1994, Proc Natl Acad Sci USA. Two-photon scanning photochemical ... Denk, W; Strickler, J.; Webb, W. (1990-04-06). "Two-photon laser scanning fluorescence microscopy". Science. 248 (4951): 73-76 ... Two-photon microscopy remains the only technique that allows the recording of activity in living brains with high spatial ... There he built one of the first super-resolution microscopes and developed a passion for scanning microscopy. He did his ...
Multiphoton excitation in laser scanning fluorescence microscopy provides for high resolution, high signal-to-noise imaging in ... Denk W, Strickler JH, Webb WW (1990). "Two-photon laser scanning fluorescence microscopy". Science. 248 (4951): 73-76. Bibcode: ... Professor Webb pioneered the techniques of Fluorescence Correlation Spectroscopy (FCS) in 1972 and Multiphoton microscopy (MPM ... In situ measurements of the dynamics of fluorescence flicker by FCS, photobleaching, phototoxicity, and induced fluorescence ...
Nie, S; Chiu, D.; Zare, R. (1994-11-11). "Probing individual molecules with confocal fluorescence microscopy". Science. ... Weiss, S. (1999-03-12). "Fluorescence Spectroscopy of Single Biomolecules". Science. American Association for the Advancement ... Zumofen, Gert; Hohlbein, Johannes; Hübner, Christian G. (2004-12-20). "Recurrence and Photon Statistics in Fluorescence ... Chung, Inhee; Bawendi, Moungi G. (2004-10-11). "Relationship between single quantum-dot intermittency and fluorescence ...
Nie, S; Chiu, D.; Zare, R. (1994). "Probing individual molecules with confocal fluorescence microscopy". Science. 266 (5187): ... Weiss, Shimon (1999). "Fluorescence Spectroscopy of Single Biomolecules". Science. 283 (5408): 1676-83. Bibcode:1999Sci... ... Chung, Inhee; Bawendi, Moungi (2004). "Relationship between single quantum-dot intermittency and fluorescence intensity decays ... Zumofen, Gert; Hohlbein, Johannes; Hübner, Christian (2004). "Recurrence and Photon Statistics in Fluorescence Fluctuation ...
Wilson, T., Optical sectioning in fluorescence microscopy. Journal of microscopy, 2011. 242: p. 111-6. Glass, Monty; Dabbs, Tim ... It generally refers to fluorescence confocal microscopy, although multi-photon microscopy and optical coherence tomography have ... Polglase, A.L., W.J. Mclaren, and S.A. Skinner, A fluorescence confocal endomicroscope for in vivo microscopy of the upper- and ... Conventional, widefield microscopy is generally unsuitable for imaging thick tissue because the images are corrupted by a ...
Fluorescence experiments involve observing fluorescent probes incorporated into the membrane. Atomic force microscopy ... Membrane fluidity can be measured with electron spin resonance, fluorescence, atomic force microscopy-based force spectroscopy ... Fluorescence correlation spectroscopy monitors the fluctuations in fluorescence intensity measured from a small number of ... Lateral motion of molecules within the membrane can be measured by a number of fluorescence techniques: fluorescence recovery ...
Such devices are used in fluorescence microscopy.[citation needed] Murphy, Douglas B.; Davidson, Michael W. (2012). ... Fundamentals of Light Microscopy and Electronic Imaging. John Wiley & Sons. p. 221.. ...
Schermelleh L (2010). "A guide to super-resolution fluorescence microscopy". J. Cell Biol. 190 (2): 165-175. doi:10.1083/jcb. ... For example, using the traditional fluorescence microscopy approach 300 - 500 factories were found but using both confocal and ... GFP polymerase fluorescence experiments have shown that the inducement of transcription in Drosophila polytene nuclei leads to ... It is possible to view them under both light and electron microscopy. The discovery of transcription factories has challenged ...
Light sheet fluorescence microscopy for the observation of development and LILBID mass spectrometry for the analysis of ... Optogenetics and light sheet fluorescence microscopy were selected as the "Method of the Year" across all fields of science and ... PMID 20601950.CS1 maint: multiple names: authors list (link) Stelzer EHK (2015). "Light-sheet fluorescence microscopy for ... Editoral (2014). "Light-sheet fluorescence microscopy can image living samples in three dimensions with relatively low ...
Nie, S; Chiu, DT; Zare, RN (11 November 1994). "Probing individual molecules with confocal fluorescence microscopy". Science. ... Early in his career, the question of whether laser-induced fluorescence (LIF) could be used to study aflatoxins spurred Zare to ... "Molecular fluorescence and photodissociation. Zare, Richard Neil". Harvard Library. Retrieved 28 March 2017. "About Richard N. ... Zare is well known for his research in laser chemistry, particularly the development of laser-induced fluorescence, which he ...
Additionally, fluorescence microscopy techniques have indicated that phagocytes also have a relatively low internal pH. Since ... Nunes P, Guido D, Demaurex N (December 2015). "Measuring Phagosome pH by Ratiometric Fluorescence Microscopy". Journal of ... This technique, which makes use of fluorescence spectroscopy, consists of adding this special dye to the cytosol of a cell. By ... the relative amount of resulting fluorescence can reveal the approximate surrounding pH. Harguindey, S; Stanciu, D; Devesa, J; ...
Linear excitation of fluorescence with confocal detection. This method is essentially confocal laser scanning microscopy. It ... In addition, fluorescence is radiated in all directions from the addressed point, so special light collection optics must be ... The reading rate is limited to 10 Mbit/s because of the fluorescence lifetime. Colossal Storage aim to develop a 3D holographic ... The intensity or wavelength of the fluorescence is different depending on whether the media has been written at that point, and ...
The name refers to the fluorescence he saw while looking at a glowing plate bombarded with X-rays.[2] ...
Fluorescence microscopy can be used to find out where the fluorescent probe is bound to the chromosomes. FISH is often used for ... Wikimedia Commons has media related to Fluorescence in situ hybridization.. *Fluorescent+in+Situ+Hybridization at the US ... Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only ... Fluorescence in situ Hybridization Photos of bacteria. *Rational design of polynucleotide probe mixes to identify particular ...
... characterization methods such as electron microscopy, X-ray diffraction, calorimetry, nuclear microscopy (HEFIB), Rutherford ... electro-fluorescence, high thermal stability, etc. ... A scanning electron microscopy image of carbon nanotubes ... A diamond cuboctahedron showing seven crystallographic planes, imaged with scanning electron microscopy ...
... the isothiocyanate derivative of fluorescein is often used to label and track cells in fluorescence microscopy applications ( ... Fluorescein is a fluorophore commonly used in microscopy, in a type of dye laser as the gain medium, in forensics and serology ... The fluorescence that is created by the dye makes problem areas more visible and easily identified. A similar concept can be ... The fluorescence of this molecule is very intense; peak excitation occurs at 494 nm and peak emission at 521 nm. ...
Advancement in microscopic techniques and technology such as fluorescence microscopy, phase-contrast microscopy, dark field ... microscopy, confocal microscopy, cytometry, transmission electron microscopy, etc. have allowed scientists to get a better idea ... Section 4. Intrinsic X-Ray Fluorescence". In Bani, Lucia (Ed.). Metallomics and the Cell. Metal Ions in Life Sciences. 12. ...
Electron microscopy. *Immunofluorescence. *Fluorescence in situ hybridization. Clinical pathology. *Clinical chemistry. * ...
Where a colorant contains fluorescence, a bi-spectral fluorescent spectrophotometer is used. There are two major setups for ... Microscopy. *Scanning electron microscope (SEM). *Transmission electron microscope (TEM). Thermochemistry. *Calorimeter * ... Traditional visible region spectrophotometers cannot detect if a colorant or the base material has fluorescence. This can make ...
Such substrates can be fabricated on a wafer scale and label-free superresolution microscopy has also been demonstrated using ... Each spectrum was specific, which is advantageous over fluorescence detection; some fluorescent markers overlap and interfere ...
Microscopy[edit]. Culture techniques will often use a microscopic examination to help in the identification of the microbe. ... quantitative PCR does not require this, as the detection system uses fluorescence and probes to detect the DNA molecules as ... Electron microscopes and fluorescence microscopes are also used for observing microbes in greater detail for research.[28] ... More detailed identification techniques involve microbial culture, microscopy, biochemical tests and genotyping. Other less ...
"for the development of super-resolved fluorescence microscopy"[108] سٹیفین ہیل رومانیا ... "for his development of crystallographic electron microscopy and his structural elucidation of biologically important nucleic ...
Single molecule probing by fluorescence and force detection. 105: 90-98. doi:10.1016/j.ymeth.2016.03.025. PMID 27038745.. ... In the case of atomic force microscopy, it may also be hard to discriminate the interaction of the tip with the studied ... optical tweezers and atomic force microscopy. The magnetic interaction is highly specific to the used superparamagnetic ... drawback of magnetic tweezers is the low temporal and spatial resolution due to the data acquisition via video-microscopy.[3] ...
Yuste R (December 2005). "Fluorescence microscopy today". Nature Methods. 2 (12): 902-4. doi:10.1038/nmeth1205-902. PMID ... Cryoelectron microscopy is used to produce lower-resolution structural information about very large protein complexes, ... Finally, the gold-standard method of cellular localization is immunoelectron microscopy. This technique also uses an antibody ... For example, indirect immunofluorescence will allow for fluorescence colocalization and demonstration of location. Fluorescent ...
Fluorescence in situ hybridization (FISH) involves fluorescent labeling of probes that bind to specific DNA sequences, used for ... While cytogenetics historically relied on microscopy to analyze chromosomes, new molecular technologies such as array ...
"for the development of super-resolved fluorescence microscopy"[115] Stefan W. Hell செருமனி. உருமேனியா[116] ... "for developing cryo-electron microscopy for the high-resolution structure determination of biomolecules in solution"[119] ... "for his development of crystallographic electron microscopy and his structural elucidation of biologically important nucleic ...
In fluorescence observations, the resolution limit of confocal microscopy is often limited by the signal to noise ratio caused ... The Development of a Modern Microscopy". Imaging & Microscopy.. online. *^ a b c Barry R. Masters: Confocal Microscopy And ... Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM), is an optical imaging technique for increasing ... Barry R. Masters: Confocal Microscopy And Multiphoton Excitation Microscopy. The Genesis of Live Cell Imaging. SPIE Press, ...
This is called Stokes Raman scattering, by analogy with the Stokes shift in fluorescence discovered by George Stokes in 1852, ... Fifth Conference on Frontiers of Aberration Corrected Electron Microscopy. 203: 60-67. doi:10.1016/j.ultramic.2018.12.006. ISSN ... "Live-Cell Bioorthogonal Chemical Imaging: Stimulated Raman Scattering Microscopy of Vibrational Probes". Accounts of Chemical ...
Energy filtered transmission electron microscopy. *Enhancement or quenching of QD, Q-wire and QW radiations ... Laser-induced fluorescence. *Laser ablation. *Laser ablation synthesis in solution. *Laser plasma acceleration ...
"DNA Base Identification by Electron Microscopy". Microscopy and microanalysis : the official journal of Microscopy Society of ... Following the development of fluorescence-based sequencing methods with a DNA sequencer,[2] DNA sequencing has become easier ... and microscopy-based techniques, such as atomic force microscopy or transmission electron microscopy that are used to identify ... Microscopy-based techniques[edit]. Main article: Transmission electron microscopy DNA sequencing. This approach directly ...
Fluorescence. *Fluorescence anisotropy. Translational Diffusion. *Analytical ultracentrifugation. *Size exclusion ...
Microscopy[edit]. Another principal tool in the diagnosis of infectious disease is microscopy. Virtually all of the culture ... A fluorescence microscope is then used to detect fluorescently labeled antibodies bound to internalized antigens within ... Microscopy may be carried out with simple instruments, such as the compound light microscope, or with instruments as complex as ... Microscopy is often also used in conjunction with biochemical staining techniques, and can be made exquisitely specific when ...
... scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), X-ray fluorescence (XRF), differential scanning ...
Electron microscopy. *Immunofluorescence. *Fluorescence in situ hybridization. Clinical pathology. *Clinical chemistry. * ...
Recognising the potentialities of laser-scanning confocal microscopy, John built a prototype microscope: with William Bradshaw ... "Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability". Nature Biotechnology. 17 (8): ... White co-developed confocal microscopy and mapped the complete nervous system of Caenorhabditis elegans, consisting of 302 ...
Thus, fluorescence light microscopy should not be used to study the glycocalyx because that particular method uses a dye. The ... Another study used cryotransmission electron microscopy and showed that the endothelial glycocalyx could be up to 11 μm thick.[ ... When vessels are stained with cationic dyes such as Alcian blue stain, transmission electron microscopy shows a small, ... The initial measurement was taken with intravital microscopy, which showed a slow-moving plasma layer, the glycocalyx, of 1 μm ...
2014 - Eric Betzig, Stefan Hell and William E. Moerner for the development of super-resolved fluorescence microscopy.[16] ... 2017 - Jacques Dubochet/ Jochiam Frank / Richard Henderson for cryo-election microscopy[19] ...
"Olympus Microscopy Resource Center. Retrieved September 19, 2007.. *^ More specifically, "blue green" 487-493 nm, "bluish green ... fluorescence, holography, machine vision, non-lethal weapons and bird control.[23] ...
Fluorescence. *Fluorescence anisotropy. Translational Diffusion. *Analytical ultracentrifugation. *Size exclusion ...
"An Introduction to Fluorescence Resonance Energy Transfer (FRET) Technology and its Application in Bioscience". BioTek ... New advancements in biotechnological experimental methods such as the use of Bessel beam plane illumination microscopy and FRET ...
These are the convergence of DNA hybridization, fluorescence microscopy, and solid surface DNA capture. The three mandatory ...
... a fact which has spurred the development of more sophisticated microscopes and fluorescence accessories. ... Fluorescence is the most rapidly expanding microscopy technique in both the medical and biological sciences, ... Advanced Techniques in Fluorescence Microscopy. * Introduction to Confocal Microscopy. Confocal microscopy offers the ability ... Fluorescence Microscopy. Section Overview: Fluorescence illumination and observation is the most rapidly expanding microscopy ...
Imaging technique based on fluorescence. Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the ... is the fluorescence lifetime, I. 0. {\displaystyle I_{0}}. is the initial fluorescence at t. =. 0. {\displaystyle t=0}. , and k ... "Fluorescence-lifetime imaging microscopy" - news · newspapers · books · scholar · JSTOR (May 2014) (Learn how and when to ... of the fluorescence signal in time bin i, the lifetime estimation is carried out by minimization of: χ. 2. =. ∑. i. [. d. i. ( ...
What is fluorescence microscopy?. Fluorescence microscopy is a non-destructive way of tracking or analyzing tissues, cells, or ... The Fluorescence Microscopy and Imaging Center provides intramural researchers with access to a variety of advanced imaging ...
Fluorescence Microscopy News and Research. RSS Fluorescence microscopy is an imaging technique used to examine cells and their ... In this interview from SfN 2018, Michelle Gal explains the fluorescence illumination systems for fluorescence microscopy, ... will display its wide range of X-Cite fluorescence illumination products for microscopy at Neuroscience 2018. ... can be detected during the examination of medical samples by means of fluorescence microscopy. ...
... fluorescence microscopy encompasses a variety of techniques that can exceed the diffraction limit - the point at which ... Super-resolution (SR) fluorescence microscopy encompasses a variety of techniques that can exceed the diffraction limit - the ... The Future of SR Microscopy. The full potential of SR microscopy is yet to be reached. The field is relatively new, and the ... Prior to the development of SR microscopy techniques, conventional wide-field microscopy was unable to distinguish ...
... Unit. The unit on main campus specializes in confocal microscopy and other advanced fluorescence ... Fluorescence Microscopy Unit. The unit on main campus specializes in confocal microscopy and other advanced fluorescence ... Fluorescence Microscopes:. With fluorescence microscopy one is able to determine localization/co-localization as well as mean ... Fluorescence Microscopes:. With fluorescence microscopy one is able to determine localization/co-localization as well as mean ...
Fluorescence microscopy Biogenesis Secretion systems Fluorescently labeled proteins Macromolecular complexes Epistasis ... Poulter NS, Pitkeathly WTE, Smith PJ, Rappoport JZ (2015) In: Verveer PJ (ed) Advanced fluorescence microscopy. Springer, New ... Zoued A., Diepold A. (2017) Defining Assembly Pathways by Fluorescence Microscopy. In: Journet L., Cascales E. (eds) Bacterial ... Johnson TL, Sikora AE, Zielke RA, Sandkvist M (2013) Fluorescence microscopy and proteomics to investigate subcellular ...
... light-sheet fluorescence microscopy conference and workshop. The program will provide a combination of virtual and in-person ... The Virtual Conference (May 9-11, 2021) will bring together innovators in microscopy from academic institutes, industry, and ... such as widefield fluorescence, confocal and multiphoton imaging. However, none of these courses are specifically designed to ...
Posted by erika pastrana , Categories: Microscopy & Imaging, Model Organisms, Neuroscience. It is now possible to map the ... Posted by Daniel Evanko , Categories: Microscopy & Imaging, Webcasts. Our very first webinar is now live. The topic is " ... Posted by Daniel Evanko , Categories: Biophysics, Editorials, Microscopy & Imaging. Light-induced damage to biological samples ... during fluorescence imaging is known to occur but receives too little attention by researchers. Read more ...
... released its new INFINITY Fluorescence Series Bundle; a fluorescence microscopy imaging solution consisting of high-end ... which gives fluorescence microscopy customers complete access to maximum frame rates, live image adjustments, and all the ... with feature-rich software packages to provide customers with a complete imaging solution for fluorescence microscopy. Each ... Lumeneras INFINITY Fluorescence Series cameras have a new sleek black enclosure, and feature Sony CCD sensors to offer high ...
To address this, we propose a content-adaptive representation of fluorescence microscopy images, the Adaptive Particle ... The APR provides a simple and efficient content-aware representation of fluosrescence microscopy images. Modern microscopes can ... Developments in fluorescence microscopy1,2,3, labeling chemistry4, and genetics5 provide the potential to capture and track ... Adaptive particle representation of fluorescence microscopy images. *Bevan L. Cheeseman ORCID: orcid.org/0000-0002-9248-51181,2 ...
Macromolecular-scale resolution in biological fluorescence microscopy. Gerald Donnert, Jan Keller, Rebecca Medda, M. Alexandra ... Macromolecular-scale resolution in biological fluorescence microscopy. Gerald Donnert, Jan Keller, Rebecca Medda, M. Alexandra ... Macromolecular-scale resolution in biological fluorescence microscopy Message Subject (Your Name) has sent you a message from ... Macromolecular-scale resolution in biological fluorescence microscopy. Gerald Donnert, Jan Keller, Rebecca Medda, M. Alexandra ...
... is an essential techniquethat allows scientists to visualise molecules (proteins, nucleic acids, ions, ... Wolf DE (2013) Fundamentals of fluorescence and fluorescence microscopy. Methods in Cell Biology 114: 69-97. ... Fluorescence microscopy is an essential technique that allows scientists to visualise molecules (proteins, nucleic acids, ions ... Keywords: fluorescence microscopy; CCD camera; confocal laser scanning; spectral analysis; optical filters; living cells ...
... www.nist.gov/programs-projects/fluorescence-microscopy-benchmarking-overview/assuring-comparability-wide-field ... and product developers are now able to implement our new procedure in quantitative wide-field fluorescence microscopy. NIST ... NIST developed a procedure for characterizing the performance of a fluorescence microscope by benchmarking the detection ... and is proven to improve the reproducibility of wide-field fluorescence imaging. Our strategy adds value to image data ...
... on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life Sciences. ... It includes a foreword by the nonlinear microscopy expert Dr. Colin Sheppard. Ed. by Karsten König ... on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life... view more ... on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life Sciences. The book ...
Ultrafast photonics for fluorescence imaging and time-resolved assays. *Multiphoton fluorescence microscopy for clinical ... Ultrafast photonics for fluorescence imaging and time-resolved assays. *Multiphoton fluorescence microscopy for clinical ... Fluorescence Microscopy. The optical microscope is one of the most widespread biophotonics tools and has allowed researchers to ... Fluorescence microscopy has enjoyed a renaissance over the last decade, partly driven by advances in light source and detector ...
The Advanced Light Microscopy facility at Science for Life laboratory cordially welcomes you to a symposium covering advanced ... applications of super-resolution fluorescence imaging from world leading experts ... Scilifelab-symposium in Advanced Fluorescence Microscopy. The Advanced Light Microscopy facility at Science for Life laboratory ... 09:50 am: Single-Molecule Localization Microscopy. Mike Heilemann - University of Frankfurt, Germany. Coffee. 11:10 am: ...
AGA(C8R)-HNG17 and the mitochondrial tracker tetramethylrhodamine methyl ester in PC-12 cells (rat pheochromocytoma, of neuronal origin) 10 min after inducing necrosis by cyanide, exhbiting co-localization of humanin and mito-tracker at the mitochondria. Both trackers are co-localized where their lifetime is the longest.
The coumarin dyes are commercially available conjugated to molecules useful in fluorescence microscopy (Molecular Probes, Inc. ... Such fields include scanned beam fluorescence microscopy, scanned beam microlithography, nanofabrication, and optical digital ... see Tsien and Waggoner, Fluorophores for Confocal Microscopy, in Handbook of Confocal Microscopy, James B. Pawley, ed., Plenum ... Fluorescence Microscopy of Living Cells in Culture, Part B, Academic Press, 1989). These techniques include placing annular and ...
BACK TO FLUORESCENCE MICROSCOPY. Questions or comments? Send us an email.. © 1998-2018 by Michael W. Davidson and The Florida ... Fluorescence Microscopy Interactive Tutorials. Photobleaching. The phenomenon of photobleaching (also commonly referred to as ... red fluorescence), respectively. Time points were taken in two-minute intervals using a fluorescence filter combination with ... The tutorial initializes with a pair of identical fluorescence images appearing in the Unbleached Image and Photobleached Image ...
This section discusses introductory concepts about fluorescence excitation and emission and the numerous applications in ... Introduction to Fluorescence - Fluorescence microscopy is a rapidly expanding and invaluable tool of investigation. Its ... Fluorescence microscopy is an excellent method of studying material that can be made to fluoresce, either in its natural form ( ... Reference Listing - The field of fluorescence spectroscopy and microscopy is experiencing a renaissance with the introduction ...
Fluorescence microscopy is almost as simple to do as bright-field microscopy, and most often it is more specific. In the past, ... Fluorescence microscopy has proved to be a useful and very cost-effective procedure for disease surveillance and for the ... Fluorescence microscopy meets all the essential requirements for laboratory examination even under constrained conditions of ... Laboratories should be more aware of the advantages of using fluorescence microscopy. The practical steps of indirect ...
... single-molecule fluorescence. TIRF microscopy images of PAmCherry1 fluorescence (a), PAGFP fluorescence (d) and the merge (g) ... The fluorescence collected during the 15,000 frames is shown in a and represents a diffraction-limited TIRF microscopy image. ( ... Photoactivatable mCherry for high-resolution two-color fluorescence microscopy.. Subach FV1, Patterson GH, Manley S, Gillette ... d-f) Magnified views of the boxed region in a are shown as TIRF microscopy (d), PALM (e) and cluster analysis images (f). PALM ...
The latest incarnation of the modern fluorescence microscope has led to a paradigm shift. This wave is about breaking the ... Conventional fluorescence microscopy uses a lens to focus a beam of light onto a spot. For example, a lens focuses the emission ... Bringing Superresolution to Fluorescence Microscopy. BioPhotonics. May 2010 Prashant Prebhat and Turan Erdogan, !%Semrock Inc ... Photoactivation localization microscopy (PALM, also referred to as F-PALM for fluorescence PALM) and stochastic optical ...
Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique for producing an image based on the differences in the ... global fluorescence lifetime imaging microscopy industry 2017fluorescence lifetime imaging microscopy market 2017global ... fluorescence lifetime imaging microscopy market shareglobal fluorescence lifetime imaging microscopy market growthglobal ... fluorescence lifetime imaging microscopy market trendsglobal fluorescence lifetime imaging microscopy market 2017 ...
Buy Protein Localization by Fluorescence Microscopy by Viki Allan from Waterstones today! Click and Collect from your local ... Protein Localization by Fluorescence Microscopy: A Practical Approach - Practical Approach Series 218 (Hardback). Viki Allan ( ... Protein Localization by Fluorescence Light Microscopy: A Practical Approach has something to offer all microscopists, giving a ... This is one major reason why fluorescence microscopy is enjoying a revival. This no-nonsense guide provides detailed, practical ...
... is an effective approach for super-resolution wide-field fluorescence microscopy with a theoretically unlimited resolution. In ... Fluorescence contrast under saturated excitation is ensured by the inherent high density of intensity minima associated with ... Compressed speckle microscopy is thus a simple approach that enables 3D super-resolved nSIM imaging with potentially ... Nonlinear structured illumination microscopy is a super-resolution technique that is challenging to extend to 3 dimensions. The ...
The technique of dual-wavelength ratio fluorescence microscopy provides a powerful tool to measure organellar pH. Unlike si.. ... Fluorescence microscopy The technique of dual-wavelength ratio fluorescence microscopy provides a powerful tool to measure ...
The base of this system is a fluorescence confocal microscope which is suitable for use in top level biological and biomedical ... The Nikon TiE is fully automated inverted widefield fluorescence microscope with builting Perfect Focus system and high-speed ... The Eclipse Ti-E inverted microscope offers improved system speed, increased flexibility and efficient multi-mode microscopy as ... Has both high resolution and flexibility to cover a wide range of experimental setups in fluorescent microscopy ...
Fluorescence Microscopy Camera by Andor Technology Ltd (Andor). With the iXon EMCCD cameras, Andor have delivered a dedicated, ... fluorescence microscopy camera microscopy microscopy camera fluorescence microscope microscope well level well camera ... No reviews were found for Andor - Model iXon EMCCD - Fluorescence Microscopy Camera. Be the first to review! ... Andor Launches iXon Life for Fluorescence Microscopy. Feb. 9, 2017. Andor Technology (Andor), an Oxford Instruments company and ...
  • Reviewed in this article are key features of fluorescence microscopy such as detecting fluorescent objects that can be faintly visible or very bright relative to the background, as well as common problems with microscope configuration. (fsu.edu)
  • The idea is to locate the specific area of interest in a specimen using the technique (phase) then, without relocating the specimen, switch the microscope to fluorescence mode. (fsu.edu)
  • Currently, many courses focus on general optical principles and the use of conventional microscope platforms, such as widefield fluorescence, confocal and multiphoton imaging. (mbl.edu)
  • NIST developed a procedure for characterizing the performance of a fluorescence microscope by benchmarking the detection threshold, saturation, and linear dynamic range to a physical artifact, such as a fluorescent material. (nist.gov)
  • The figure shows the most common form of fluorescence microscope where excitation light is reflected by a dichroic beamsplitter and illuminates the field of view to produce fluorescence at a wavelength that is transmitted by the dichroic beamsplitter. (imperial.ac.uk)
  • The fluorescence microscope was devised in the early part of the twentieth century by August K hler, Carl Reichert, and Heinrich Lehmann, among others. (fsu.edu)
  • What is more, the fluorescence microscope can reveal the presence of fluorescing material with exquisite sensitivity. (fsu.edu)
  • Although the fluorescence microscope cannot provide spatial resolution below the diffraction limit of the respective specimens, the presence of fluorescing molecules below such limits is made remarkably visible. (fsu.edu)
  • More recently, less expensive fluorescence microscopes have been developed, and accessories are now available that convert a bright-field microscope into a fluorescence microscope. (who.int)
  • The latest incarnation of the modern fluorescence microscope has led to a paradigm shift. (photonics.com)
  • So-called super resolution microscopy allows for the visualization of cellular samples with a resolution similar to that of an electron microscope, yet it retains the advantages of an optical fluorescence microscope. (photonics.com)
  • So what is wrong with a conventional fluorescence microscope? (photonics.com)
  • The base of this system is a fluorescence confocal microscope which is suitable for use in top level biological and biomedical research and surface analysis in material science applications. (dcu.ie)
  • The Nikon TiE is fully automated inverted widefield fluorescence microscope with builting Perfect Focus system and high-speed motorization. (dcu.ie)
  • The Eclipse Ti-E inverted microscope offers improved system speed, increased flexibility and efficient multi-mode microscopy as part of a fully-integrated microscope system that is ideal for high-end research and with its environmental control, it is perfect for live cell imaging. (dcu.ie)
  • The fluorescence microscope (wide‐field, scanning, confocal, one‐photon excitation, multiphoton excitation) is an extremely useful and ubiquitous instrument in biological and medical laboratories. (els.net)
  • The fluorescence microscope provides enhanced contrast, single protein specificity and single molecule sensitivity. (els.net)
  • Progress in the technical development over more than 100 years of instrument design and fluorescent probe synthesis has contributed to the continuing widespread utility of the fluorescence microscope. (els.net)
  • Recently, genetically expressed fluorescent proteins and quantum dots provide new research capabilities for the intravital fluorescence microscope. (els.net)
  • The reader will gain an understanding of the development of both the fluorescence microscope instrumentation and the field of fluorescent probe synthesis, development, limitations and applications to the life sciences. (els.net)
  • The fluorescence microscope can provide single molecule detection sensitivity. (els.net)
  • The fluorescence microscope can provide single protein selectivity. (els.net)
  • A major advance in fluorescence microscope is the development of multiphoton excitation microscopy. (els.net)
  • Development of a confocal laser scanning fluorescence microscope using two. (spiedigitallibrary.org)
  • Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. (open.ac.uk)
  • We present the first realization of a Two-Color Two-Photon Laser-Scanning Microscope (2c2pLSM) and UV fluorescence images of cells acquired with this technique. (biomedsearch.com)
  • This technique for Laser-Scanning Microscopy extends the excitation wavelength range of a Ti:Sa powered fluorescence microscope to the UV. (biomedsearch.com)
  • For the purpose of this report, fluorescence microscopy or microscope is defined as an optical microscope that uses fluorescence and phosphorescence instead of, or in addition to, reflection and absorption, to study properties of organic or inorganic substances. (bccresearch.com)
  • A fluorescence microscope is based on the phenomenon that certain materials emit energy which is detectable as visible light when irradiated with the light of a specific wavelength. (bccresearch.com)
  • A fluorescence microscope uses a much more intense light source that excites a fluorescent species in a sample of interest. (bccresearch.com)
  • Bruker has developed a fully integrated fluorescence microscope for TriboIndenter Series Instruments. (bruker.com)
  • The fluorescence microscope utilizes user-definable filter sets for use with a wide range of fluorescent dyes and is also capable of bright field imaging, allowing the nanoindenter system to be effectively utilized in a variety of research areas. (bruker.com)
  • We describe a method of obtaining optical sectioning with a standard wide-field fluorescence microscope. (nih.gov)
  • This book provides a comprehensive account of the theory of image formation in a confocal fluorescence microscope as well as a practical guideline to the operation of the instrument, its limitations, and the interpretation of confocal microscopy data. (spie.org)
  • Since its introduction in the late seventies, the confocal fluorescence microscope has advanced rapidly from a complex instrument that could be used by specialists only, to a commercial product, which is part of the standard repertoire of modern biological research. (spie.org)
  • Microscope Zeiss Elyra PS.1 use a technologies of structured illumination (SIM) and precise localization (PALM/STORM) to overcome so called diffraction barrier, which limits us to resolution of cca 200 nm in conventional light microscopy. (cuni.cz)
  • Microscope can use a technology of TIRF (Total Internal Reflection Fluorescence) to increase signal to noise ratio in the space near to cover glass. (cuni.cz)
  • This unit discusses the optics and other components of the fluorescence microscope, their functions and adjustments, and the digital darkroom. (currentprotocols.com)
  • A technique using a light microscope to study properties of organic or inorganic substances using the phenomena of fluorescence and phosphorescence instead of, or in addition to, reflection and absorption . (biology-online.org)
  • This fluorescence tutorial explains the optical elements in the light path and the operating mode of fluorescence microscopy taking the example of an inverted microscope which can be used for transmitted light contrasting methods and fluorescence microscopy. (leica-microsystems.com)
  • Diagram illustrating the optical filters and light path of a fluorescence microscope. (azooptics.com)
  • The light path through lenses and sample in basic brightfield microscopy (A). Antique 19th century drum-style compound microscope (B). (thermofisher.com)
  • Accessories for X-Cite fluorescence illuminators including optical power measurement, lamp replacements, liquid light guides, and microscope adapters for Zeiss, Olympus, Leica and Nikon. (microscopeworld.com)
  • The design is based on an apertured fluorescence detector placed close to the back aperture of the objective lens and can be easily incorporated into virtually any confocal microscope. (osapublishing.org)
  • Multiple-view imaging was developed as an extension to the light-sheet based fluorescence microscope (LSFM), a novel technique that seems to be better suited for multiple-view imaging than any other fluorescence microscopy method to date. (uni-heidelberg.de)
  • Fluorescence illumination and observation is the most rapidly expanding microscopy technique employed today, both in the medical and biological sciences, a fact which has spurred the development of more sophisticated microscopes and numerous fluorescence accessories. (fsu.edu)
  • Unfortunately, fluorescence microscopes do not directly output the shapes and locations of objects through time. (nature.com)
  • This monograph focuses on modern femtosecond laser microscopes for two photon imaging and nanoprocessing, on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life. (eurekalert.org)
  • In the past, the high cost of fluorescence microscopes has prevented the wider application of this method. (who.int)
  • SRRF-Stream unlocks the means to perform real time super resolution microscopy on conventional modern fluorescence microscopes. (environmental-expert.com)
  • The basic principle of fluorescence imaging is the same for all microscopes: The sample is first chemically labelled with fluorescent molecules called fluorophores. (mpg.de)
  • The market is broken down by illumination methods, types of fluorescence microscopes, technology, region, and applications (end users). (bccresearch.com)
  • The new 2.0 version of the Basler Microscopy Software also offers exposure compensation and a new zoom feature for stereo microscopes. (baslerweb.com)
  • Fluorescence microscopes are equipped to observe the fluorescence of one or more specific fluorophores to localize specific molecules and analyzed cellular structures. (currentprotocols.com)
  • Leica Microsystems' fluorescence stereo microscopes use the TripleBeam technology , a separate (third) beam path for fluorescence illumination of the sample without a dichroic mirror. (leica-microsystems.com)
  • Fluorescence microscopes and imaging systems make use of fluorescent biomarkers and fluorescence filter sets to produce bright and high-contrast images of biomolecules, organelles, cells, tissues, organs, and organ systems. (azooptics.com)
  • Image quality is highly dependent on the design and the grade of performance seen in these fluorescence filters that are integrated into fluorescence microscopes. (azooptics.com)
  • Fluorescence microscopes, imaging systems, flow cytometers, and DNA sequencers all use a light source, whether that is a laser, LED, or broadband lamp, in order to excite the fluorescent tags within a sample. (azooptics.com)
  • Three types of optical filters are needed for epi-fluorescence and other standard fluorescence microscopes to function properly. (azooptics.com)
  • Learn the basic physics behind the excitation and emission of a fluorophore and how microscopes channel and capture the light generated to make a fluorescence-based image. (thermofisher.com)
  • When the technique of fluorescence resonance energy transfer ( FRET ) is applied to optical microscopy, it permits determination of the approach between two molecules within several nanometers. (fsu.edu)
  • FLIM has been used to image interactions between proteins such as receptor oligomerisation and to reveal protein phosphorylation by detecting fluorescence resonance energy transfer (FRET). (rsc.org)
  • Fluorescence Resonance Energy Transfer Microscopy for Measuring Chromatin Complex Structure and Dynamics. (nih.gov)
  • There is a chamber with CO 2 atmosphere for live cell imaging available.It is possible to do multicolor analysis as well as recording of physiologial processes, 3D reconstructions, FRAP (Fluorescence Recovery After Photobleaching) and FRET (Fluorescence Resonance Energy Transfer) analysis an tile scans. (cuni.cz)
  • Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer. (currentprotocols.com)
  • Resonance energy transfer microscopy. (currentprotocols.com)
  • Membranes of living cells are characterized by laser-assisted fluorescence microscopy , in particular a combination of microspectrofluorometry , total internal reflection fluorescence microscopy ( TIRFM ), fluorescence lifetime imaging ( FLIM ) and Förster resonance energy transfer (FRET) spectroscopy . (rsc.org)
  • The processes, such as fluorescence resonance energy transfer (FRET) and stimulated emission depletion (STED), are used in microscopy. (azooptics.com)
  • These techniques include FRET (fluorescence resonance energy transfer), FLIM (fluorescence lifetime imaging microscopy), FCS (fluorescence correlation spectroscopy), and FRAP (fluorescence recovery after photobleaching). (picoquant.com)
  • Super-resolution fluorescence microscopy has proven to be a fundamental tool in biology to unveil processes occurring at the nano-scale 1 . (nature.com)
  • This limit was regarded as insurmountable − until researchers developed super-resolution fluorescence microscopy methods, such as STED and PALM / STORM, within the past two decades. (mpg.de)
  • With the help of super-resolution fluorescence microscopy , it is even possible to image sub-resolution structures. (leica-microsystems.com)
  • Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy. (sigmaaldrich.com)
  • A method to fluorescently stain the surfaces of both Gram-negative and Gram-positive bacterial cells compatible with super-resolution fluorescence microscopy is presented. (sigmaaldrich.com)
  • Unlike other modes of optical microscopy based on macroscopic specimen features, such as birefringence, fluorescence microscopy is capable of imaging the distribution of a single molecular species based solely on the properties of fluorescence emission. (fsu.edu)
  • Widefield fluorescence and laser scanning confocal microscopy rely on secondary fluorescence emission as an imaging mode, primarily due to the high degree of sensitivity afforded by the techniques. (fsu.edu)
  • Optical highlighters generally display little or no initial fluorescence under excitation at the imaging wavelength, but increase their fluorescence intensity after activation by irradiation at a different wavelength. (fsu.edu)
  • The lasers employed in optical microscopy are high-intensity monochromatic light sources, which are useful for many techniques including optical trapping, lifetime imaging studies, and photobleaching recovery. (fsu.edu)
  • Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the differences in the exponential decay rate of the fluorophore from a sample. (wikipedia.org)
  • It can be used as an imaging technique in confocal microscopy , two-photon excitation microscopy , and multiphoton tomography. (wikipedia.org)
  • which allows one to view contrast between materials with different fluorescence decay rates (even if those materials fluoresce at exactly the same wavelength), and also produces images which show changes in other decay pathways, such as in FRET imaging . (wikipedia.org)
  • Fluorescence microscopy is an imaging technique used to examine cells and their internal components. (news-medical.net)
  • With conventional wide-field microscopy all markers are switched on at the same time, causing overlapping and blurred imaging to be produced. (news-medical.net)
  • In comparison to other microscopy techniques, STED is able to produce video-rate imaging and works well with a variety of markers. (news-medical.net)
  • Our latest addition, the AMNIS imaging flow cytometer combines the best of microscopy and flow cytometry, providing a very fast analyses tool to image thousands of cells or particles in suspension for further analyses. (sun.ac.za)
  • Light-induced damage to biological samples during fluorescence imaging is known to occur but receives too little attention by researchers. (nature.com)
  • a fluorescence microscopy imaging solution consisting of high-end scientific cameras, feature-rich software packages, and a 5-year warranty. (prweb.com)
  • The INFINITY Fluorescence Series Bundle includes an INFINITY3 series camera, with feature-rich software packages to provide customers with a complete imaging solution for fluorescence microscopy. (prweb.com)
  • Lumenera's INFINITY3 series cameras have proven to excel in fluorescence imaging - even under challenging conditions," said Eric Ramsden, Product Manager at Lumenera Corporation. (prweb.com)
  • Using multiple different imaging modalities, scientists have adapted fluorescence microscopy to advance our knowledge in all areas of biology and across length scales that range from tens of millimetres to a few nanometres. (els.net)
  • Fluorescence imaging systems are built to excite molecules and then collect the emitted photons. (els.net)
  • Imaging resolution has been limited to a diffraction limit based on the nature of our ability to collect fluorescence light through the objective, but new techniques have allowed for imaging to almost arbitrary precision. (els.net)
  • This method has already been implemented by several universities and companies, and is proven to improve the reproducibility of wide-field fluorescence imaging . (nist.gov)
  • Indeed, for much biological research, "microscopy" has become almost synonymous with "imaging", although many concepts developed for microscopy are being translated to optical imaging at larger scales. (imperial.ac.uk)
  • For studies of disease mechanisms, microscopy is increasingly evolving from imaging thin layers of cells on a coverslip to studies of 3-D cell cultures, which provide a better representation of the in vivo context, and to imaging of cellular processes in live organisms - from worms and fruit flies to humans. (imperial.ac.uk)
  • Lack of green fluorescence and single-molecule behavior make monomeric PAmCherry1 a preferred tag for two-color diffraction-limited photoactivation imaging and for super-resolution techniques such as one- and two-color photoactivated localization microscopy (PALM). (nih.gov)
  • Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique for producing an image based on the differences in the exponential decay rate of the fluorescence from a fluorescent sample. (mynewsdesk.com)
  • This report focuses on the Fluorescence Lifetime Imaging Microscopy in Global market, especially in North America, Europe and Asia-Pacific, South America, Middle East and Africa. (mynewsdesk.com)
  • There are 15 Chapters to deeply display the global Fluorescence Lifetime Imaging Microscopy market. (mynewsdesk.com)
  • Fluorescence lifetime imaging ( FLIM ) can report on photophysical events that are difficult or impossible to observe by fluorescence intensity imaging, and time-resolved fluorescence anisotropy imaging (TR-FAIM) can measure the rotational mobility of a fluorophore in its environment. (rsc.org)
  • Compressed speckle microscopy is thus a simple approach that enables 3D super-resolved nSIM imaging with potentially considerably reduced acquisition time and photobleaching. (nature.com)
  • Here, we demonstrate the possibility to achieve 3D super-resolution imaging by a single 2D raster scan under saturated fluorescence excitation with tightly focused speckle patterns. (nature.com)
  • Andor Technology (Andor), an Oxford Instruments company and world leader in scientific imaging and spectroscopy solutions, today announced the launch of the new ultrasensitive iXon Life Electron Multiplying CCD (EMCCD) camera platform, exclusively for fluorescence microscopy. (environmental-expert.com)
  • Apex Market Reports, recently published a detailed market research study focused on the "Fluorescence Lifetime Imaging Microscopy Market" across the global, regional and country level. (openpr.com)
  • The report provides 360° analysis of "Fluorescence Lifetime Imaging Microscopy Market" from view of manufacturers, regions, product types and end industries. (openpr.com)
  • The research report analyses and provides the historical data along with current performance of the global PP Pipe industry, and estimates the future trend of Fluorescence Lifetime Imaging Microscopy on the basis of this detailed study. (openpr.com)
  • The study shares "Fluorescence Lifetime Imaging Microscopy" performance both in terms of volume and revenue. (openpr.com)
  • The market research report explores the Fluorescence Lifetime Imaging Microscopy Market across the globe along with major regions and countries. (openpr.com)
  • Moreover, the research study classifies the Fluorescence Lifetime Imaging Microscopy Market based on major product types, application and end users industries of Fluorescence Lifetime Imaging Microscopy. (openpr.com)
  • Besides, the report also covers geographical segmentation for Fluorescence Lifetime Imaging Microscopy Market. (openpr.com)
  • The competitive landscape of the global market for Fluorescence Lifetime Imaging Microscopy is determined by assessing the major industry participants, production capacity, production capacity utilization rate, Fluorescence Lifetime Imaging Microscopy Market's production chain, pricing by each manufacturer and the revenue generated by each manufacturer in the Fluorescence Lifetime Imaging Microscopy Market globally. (openpr.com)
  • The Global Fluorescence Lifetime Imaging Microscopy Market 2018 is further analyzed on the basis of product pricing, Fluorescence Lifetime Imaging Microscopy production volume, data pertaining to demand and Fluorescence Lifetime Imaging Microscopy supply, and the revenue garnered by the product. (openpr.com)
  • The report provides upstream and downstream analysis covering major raw material used in manufacturing of Fluorescence Lifetime Imaging Microscopy along with detailed manufacturing sources. (openpr.com)
  • Various methodical tools such as investment returns, feasibility, SWOT analysis and market attractiveness analysis has been implemented in the research study to present a comprehensive, detailed study of the industry for Fluorescence Lifetime Imaging Microscopy across the world. (openpr.com)
  • 2002) Biomedical applications of fluorescence lifetime imaging. (els.net)
  • 2008) Label‐free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy. (els.net)
  • Fluorescence microscopy imaging has rapidly evolved in these years and provided new means for studying microbial behavior, cell motion, or intracellular processes in vivo. (hindawi.com)
  • The fluorescence image signal features a third-order dependence on the excitation power, also providing intrinsic 3-D imaging. (spiedigitallibrary.org)
  • The central obstacle hindering progress in this area is the shortage of methods for labeling proteins for imaging by both fluorescence microscopy and electron microscopy. (pnas.org)
  • The key finding is that VIPER delivers a variety of chemical reporters, thus enabling effortless switching from fluorescence microscopy to high-resolution electron microscopy imaging without changing the genetically encoded tag. (pnas.org)
  • It was the summer of 2005 when in the laboratory of Fred Wouters we started the first autonomous runs of our new in-house developed high throughput fluorescence lifetime imaging (FLIM) system. (mdpi.com)
  • A new method for fluorescence imaging has been developed that can obtain spatial distributions of large numbers of fluorescent molecules on length scales shorter than the classical diffraction limit. (nih.gov)
  • We characterize the structure and mechanical properties of 1,3,5-benzenetricarboxamide (BTA) supramolecular polymers using correlative AFM and fluorescence imaging. (tue.nl)
  • In addition to the known advantages of multi-photon microscopy like intrinsic 3D resolution, reduced photo damage and high penetration depth 2c2pLSM offers the possibility of using standard high numeric aperture objectives for UV fluorescence imaging. (biomedsearch.com)
  • Hence, it is an ideal tool for label free fluorescence studies and imaging of intrinsic protein fluorescence which originates mainly from tryptophan. (biomedsearch.com)
  • Nanoindentation and fluorescence imaging is combined utilizing a high-precision automated staging system and provides exceptional spatial synchronicity between the techniques. (bruker.com)
  • Our PowerPack portfolio now features two cameras particularly suitable for fluorescence imaging: the Microscopy ace 2.3 MP Mono offers a resolution of 2.3 MP combined with high sensitivity thanks to its large pixel size. (baslerweb.com)
  • To make fluorescence imaging more convenient and to save the user´s time, the software has also been enhanced with color presets for the most common fluorescence markers. (baslerweb.com)
  • It is possible to do one channel FLIM (Fluorescence Life-time IMaging), FCS (Fluorescence Correlation Spectroscopy) and RICS (Raster Image Correlation Microscopy) measurents which allow us uncover molecular relationships and functions. (cuni.cz)
  • Three‐dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two‐photon excitation laser scanning microscopy. (currentprotocols.com)
  • Fluorescence imaging creates a window on the cell. (currentprotocols.com)
  • This one week, intensive microscopy course will cover all aspects of the technology from the principals of fluorescence imaging to multidimensional imaging in living cells. (mdibl.org)
  • The THUNDER Imager Tissue allows real-time fluorescence imaging of 3D tissue sections typically used in neuroscience and histology research. (leica-microsystems.com)
  • Göttfert F, Pleiner T, Heine J, Westphal V, Görlich D, Sahl SJ, Hell SW (2017) Strong signal increase in STED fluorescence microscopy by imaging regions of subdiffraction extent. (springer.com)
  • Now you can take the Molecular Probes School of Fluorescence Basics of Imaging: Chapter 1-Fundamentals of fluorescence microscopy with you on your devices. (thermofisher.com)
  • It's pretty important to understand the difference between magnification and resolution when it comes to getting a good result when you're doing fluorescence imaging. (thermofisher.com)
  • This we're going to be talking about life cell imaging and fluorescence microscopy. (coursera.org)
  • makes fluorescence microscopy compatible with live cell imaging over a time course. (coursera.org)
  • This webinar is an ideal opportunity for academics and researchers in cell and development biology fields to see how light-sheet microscopy can benefit their research and imaging potential. (bruker.com)
  • Light-sheet fluorescence microscopy is opening the door for many researchers to perform high-speed 3D imaging of large and delicate samples while still being able observe fast subcellular processes and interactions within the organ, organoid or organism context. (bruker.com)
  • Watch this webinar to gain insights into the basics of light-sheet microscopy and the critical details that produce the best light-sheet imaging results. (bruker.com)
  • Super Resolution imaging using single molecule localization techniques (PALM, STORM) provides the highest resolution available for a fluorescence microscopy modality. (bruker.com)
  • Is the Subject Area "Fluorescence imaging" applicable to this article? (plos.org)
  • With Quant-View technology-which quantifies imaging data in photoelectrons-disabled (panels A to C), reported fluorescence signal increased with greater applied EM Gain. (bioopticsworld.com)
  • In this contribution, the LSFM properties, which are important for multiple-view imaging, are characterized and the implementation of LSFM based multiple-view microscopy is described. (uni-heidelberg.de)
  • In recent years, non-linear imaging techniques such as multiphoton fluorescence and harmonic generation microscopy have been successfully applied to dermatological imaging. (spie.org)
  • We present a multimodal visible light optical coherence microscopy (OCM) and fluorescence imaging (FI) setup. (spiedigitallibrary.org)
  • 1 The use of OCT or optical coherence microscopy (OCM) also expanded into a wide range of application fields such as neuro-, skin, and endoscopic imaging. (spiedigitallibrary.org)
  • One promising possibility is to combine OCT and fluorescence imaging (FI). (spiedigitallibrary.org)
  • AFM and fluorescence imaging with specific lectin probes demonstrated that the polarized surface structure correlates with a heterogeneous distribution of WTAs, the latter being absent from the surface of the poles. (wur.nl)
  • Professor Hell received the Nobel Prize in Chemistry in 2014 'for the development of super-resolved fluorescence microscopy,' and the Kavli Prize in nanoscience that same year for his 'transformative contributions to the field of nano-optics that have broken long-held beliefs about the limitations of the resolution limits of optical microscopy and imaging. (carnegiescience.edu)
  • Here we present a correlative scanning TEM (STEM) and fluorescence microscopy technique for imaging viable cells of Magnetospirillum magneticum strain AMB-1 in liquid using an in situ fluid cell TEM holder. (osti.gov)
  • Fluorescently labeled cells were immobilized on microchip window surfaces and visualized in a fluid cell with STEM, followed by correlative fluorescence imaging to verify their membrane integrity. (osti.gov)
  • Notably, the post-STEM fluorescence imaging indicated that the bacterial cell wall membrane did not sustain radiation damage during STEM imaging at low electron dose conditions. (osti.gov)
  • We compare different FLIM methods: a chief advantage of wide-field time-gating and phase modulation methods is the speed of acquisition whereas for time-correlated single photon counting ( TCSPC ) based confocal scanning it is accuracy in the fluorescence decay. (rsc.org)
  • The manuals focus lies on the core F-techniques FRET, FLIM, FCS, and FRAP, but recognizes recent developments that lead to many interesting variants currently in use, such as fluorescence cross correlation spectroscopy (FCCS). (picoquant.com)
  • Learn the basic concepts of fluorescence, a member of the ubiquitous luminescence family of processes in which susceptible molecules emit light from electronically excited states created by either a physical, mechanical, or chemical mechanism. (fsu.edu)
  • The CAF Fluorescence Microscopy Unit houses fluorescence-based instruments for assessing the various properties of cells, particles or molecules of interest in a wide variety of samples. (sun.ac.za)
  • Fluorescence microscopy is an essential technique that allows scientists to visualise molecules (proteins, nucleic acids, ions, metabolites, carbohydrates and lipids), large structures and whole cells in fixed and living specimens as well as single molecules, assemblies and enzymes in vitro . (els.net)
  • Fluorescence is an intrinsic property of some molecules and proteins that causes them to absorb and then emit light at given wavelengths. (els.net)
  • Localisation of molecules is only one of many readouts that scientists can obtain from fluorescence microscopy. (els.net)
  • Increasingly, however, fluorescence instrumentation aims to provide more information than just the distribution of specific fluorescent molecules. (imperial.ac.uk)
  • Recently, it was realized that ToF sensors could also be used for the detection of fluorescence and of the minute changes in the nanosecond-lived electronic states of fluorescent molecules. (mdpi.com)
  • The fluorescence is imaged by a CCD camera, and then the molecules are either reversibly inactivated or irreversibly photobleached to remove them from the field of view. (nih.gov)
  • FPALM images are compared with images of the same molecules by widefield fluorescence. (nih.gov)
  • Hence, SAF Microscopy is ideally suited to discriminate between molecules and particles at or close to surfaces and all other specimen present in the bulk. (wikipedia.org)
  • Absorption of energy as light by some molecules and emission of energy as fluorescence can occur only at certain wavelengths, which are characteristic for a given molecule (fluorophore). (currentprotocols.com)
  • many molecules that we're looking at the fluorescence of. (coursera.org)
  • The new approach, called dispersion-relation fluorescence spectroscopy (DFS), labels molecules of interest with a fluorophore whose motion, the researchers write, "gives rise to spontaneous fluorescence intensity fluctuations that are analyzed to quantify the governing mass transport dynamics. (phys.org)
  • I thought you might be interested in this item at http://www.worldcat.org/oclc/76213907 Title: Cell substrate distance measurement in correlation with distribution of adhesion molecules by fluorescence microscopy Author: Yoriko Iwanaga Publisher: [S.l.] : [s.n.], 2000. (worldcat.org)
  • In order to generate enough excitation light intensity to furnish secondary fluorescence emission capable of detection, powerful light sources are needed such as mercury or xenon arc (burner) lamps. (fsu.edu)
  • To observe fluorescence, one of these pathways must be by spontaneous emission of a photon. (wikipedia.org)
  • The 10- to 12-fold multilateral increase in resolution below the diffraction barrier has been enabled by the elimination of molecular triplet state excitation as a major source of photobleaching of a number of dyes in stimulated emission depletion microscopy. (pnas.org)
  • Allowing for relaxation of the triplet state between subsequent excitation-depletion cycles yields an up to 30-fold increase in total fluorescence signal as compared with reported stimulated emission depletion illumination schemes. (pnas.org)
  • d) An example of a triple bandpass excitation filter, dichroic beamsplitter and emission filter set that can be used in a filter cube, coupled with separate emission and excitation filters to perform three‐colour fluorescence images. (els.net)
  • Often fluorescence signals are analysed spectroscopically, e.g. in terms of excitation or emission wavelength, fluorescence lifetime or polarisation, to provide information on the local fluorophore environment, to study interactions of biomolecules and to distinguish different contributions from complex mixtures of fluorophores- as often occur in the autofluorescence of unlabelled biological tissue. (imperial.ac.uk)
  • Because photobleaching, a very poorly understood phenomenon, leads to a dramatic loss of fluorescence emission intensity in most specimens, controlling the artifact is critical in order to successfully capture satisfactory images. (fsu.edu)
  • Time points were taken in two-minute intervals using a fluorescence filter combination with bandwidths tuned to excite the three fluorophores simultaneously while also recording the combined emission signals. (fsu.edu)
  • To achieve maximum fluorescence intensity, the fluorochrome is usually excited at the wavelength at the peak of the excitation curve, and the emission is selected at the peak wavelength (or other wavelengths chosen by the observer) of the emission curve. (fsu.edu)
  • In fluorescence microscopy , the fluorescence emission can be characterised not only by intensity and position, but also by lifetime, polarization and wavelength. (rsc.org)
  • Second, speckles exhibit strong intensity contrasts since naturally containing a high density of optical vortices of topological charge one 27 , such as typically used in STimulated Emission Depletion (STED) microscopy 28 . (nature.com)
  • Here the cell structures are labelled with fluorescent dyes that respond with fluorescence emission upon stimulation by light of a certain wavelength. (uni-protokolle.de)
  • Hell SW and Wichmann J (1994) Breaking the diffraction resolution limit by stimulated emission: stimulated‐emission‐depletion fluorescence microscopy. (els.net)
  • The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation. (sciencemag.org)
  • Fluorescence image was taken at blue excitation light and green emission. (bruker.com)
  • For example, some fluorescence systems have been designed to detect the emission signal from a single molecule. (azooptics.com)
  • Structured illumination microscopy (SIM) images of a bovine pulmonary artery endothelial cell stained with fluorescent dyes for mitochondria, actin, nucleus. (news-medical.net)
  • Nonlinear structured illumination microscopy (nSIM) is an effective approach for super-resolution wide-field fluorescence microscopy with a theoretically unlimited resolution. (nature.com)
  • Typically, the fragility of dyes has never allowed recording several transverse planes in saturated-excitation structured-illumination microscopy. (nature.com)
  • Gustafsson MG (2000) Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. (springer.com)
  • Using structured illumination microscopy and stochastic optical reconstruction microscopy, which require high quantum yield or specialized dyes, we show that this staining method may be used to resolve the bacterial cell surface with sub-diffraction-limited resolution. (sigmaaldrich.com)
  • RESOLFT (reversible saturable/switchable optically linear fluorescence transition) is a generalized name for STED or SPEM. (photonics.com)
  • FPALM images of PA-GFP on a terraced sapphire crystal surface were compared with atomic force microscopy and show that the full width at half-maximum of features approximately 86 +/- 4 nm is significantly better than the expected diffraction-limited optical resolution. (nih.gov)
  • Here, we used single-molecule atomic force microscopy (AFM) combined with fluorescence microscopy to image the distribution of wall teichoic acids (WTAs) in. (wur.nl)
  • When a population of fluorophores is excited by an ultrashort or delta pulse of light, the time-resolved fluorescence will decay exponentially as described above. (wikipedia.org)
  • By labelling different proteins with fluorophores emitting at different wavelengths, spectrally-resolved fluorescence images can be used to provide information about temporal and spatial colocalisation of the different labelled proteins, which can provide insights into molecular processes. (imperial.ac.uk)
  • The tutorial is designed to randomly alter photobleaching rates for individual fluorophores upon reloading, so that clicking on the Shutter button repeatedly will demonstrate how the image appears as the fluorescence bleaching rate changes for each of the color signals. (fsu.edu)
  • Fluorescence photoactivation localization microscopy (FPALM) analyzes thousands of single fluorophores per acquisition, localizing small numbers of them at a time, at low excitation intensity. (nih.gov)
  • The excitation of fluorophores having single-photon absorption in the ultraviolet with a stream of strongly focused subpicosecond pulses of red laser light has made possible fluorescence images of living cells and other microscopic objects. (sciencemag.org)
  • In super-resolution microscopy, however, fluorophores that preferentially exist in a non-fluorescent state, but occasionally re-arrange into a fluorescent form, can give better results. (epfl.ch)
  • Fluorophores, also called fluorochromes, are generally present in samples that have been studied with fluorescence. (azooptics.com)
  • Notes: SSIM is also referred to as SPEM (saturated pattern excitation microscopy). (photonics.com)
  • Recently, different methods have been reported to overcome this limit, among them stochastic optical reconstruction microscopy (STORM). (uni-protokolle.de)
  • With fluorescence microscopy one is able to determine localization/co-localization as well as mean intensity of a molecule of interest and with some advancements in the techniques structural analysis on a nanoscale is now possible. (sun.ac.za)
  • Johnson TL, Sikora AE, Zielke RA, Sandkvist M (2013) Fluorescence microscopy and proteomics to investigate subcellular localization, assembly, and function of the type II secretion system. (springer.com)
  • Protein Localization by Fluorescence Light Microscopy: A Practical Approach has something to offer all microscopists, giving a solid grounding to the novice whilst extending the range of the experienced user. (waterstones.com)
  • Goedhart & Gadella (2009) show methods of microscopy that quantify the localization, conformation and activity of key signalling proteins in living cells. (royalsocietypublishing.org)
  • Nanoscopy allow us observe structures with 2-3x higher resolution with SIM and up to 5 nm with localization microscopy. (cuni.cz)
  • Fluorescence microscopy even allows users to determine the distribution of a single molecule species, its amount and its localization inside a cell. (leica-microsystems.com)
  • See how single molecule localization microscopy can be used to image chromosomal regions and visualize chromosomal structure. (bruker.com)
  • Single molecule localization microscopy allows for the direct investigation of the molecular positions and distribution of proteins within the cellular environment, creating data sets that can be analyzed conveniently through numerous statistical analysis methods. (bruker.com)
  • Fluorescence lifetime depends on the local micro-environment of the fluorophore, thus precluding any erroneous measurements in fluorescence intensity due to change in brightness of the light source, background light intensity or limited photo-bleaching. (wikipedia.org)
  • This technology allows for the rapid and concurrent measurement of the physical and fluorescent parameters of 0.5-50µm particles, as they pass through a laser beam, giving information about the relative size, internal complexity and fluorescence intensity of the particles or cells of interest. (sun.ac.za)
  • As in fluorescence microscopy, the information in different areas of a scene is not encoded in absolute intensity differences, but in relative differences compared to the local brightness. (nature.com)
  • Fluorescence contrast under saturated excitation is ensured by the inherent high density of intensity minima associated with optical vortices in polarized speckle patterns. (nature.com)
  • The rate of photobleaching is controlled by the intensity of the laser used to excite the fluorescence, in this case an Ar+ ion laser. (nih.gov)
  • They derived a relationship between the observed fluorescence intensity and the distance of the fluorophore from a reflective silicon surface. (wikipedia.org)
  • Laboratories should be more aware of the advantages of using fluorescence microscopy. (who.int)
  • Researchers and product developers are now able to implement our new procedure in quantitative wide-field fluorescence microscopy . (nist.gov)
  • The global fluorescence microscopy market should reach $768.4 million by 2023 from $547.8 million in 2018 at a compound annual growth rate (CAGR) of 7.0% for the period of 2018 to 2023. (bccresearch.com)
  • The scope of this report covers the global fluorescence microscopy market for various end-user application industries. (bccresearch.com)
  • Further, it explains the major drivers, regional dynamics of the global fluorescence microscopy market and current trends within the industry. (bccresearch.com)
  • The report covers hardware revenue of companies active in the global fluorescence microscopy market. (bccresearch.com)
  • The report begins with an introduction to the evolving global fluorescence microscopy market and the various factors which impact the market. (bccresearch.com)
  • Vahid Sandoghdar and his colleagues have been working on alternative methods to increase the performance of high-resolution fluorescence microscopy for several years now − with considerable success, as the Erlangen physicist reports: "Our new approach allows us to display structures that are separated by a distance of less than five angstroms, i.e. half a nanometre. (mpg.de)
  • Fluorescence microscopy can be combined with contrast enhancing techniques such as DIC illumination to minimize the effects of photobleaching by locating a specific area of interest in a specimen using DIC . (fsu.edu)
  • To minimize photobleaching, fluorescence microscopy can be combined with phase contrast illumination. (fsu.edu)
  • b) In a FRAP experiment, a region is photobleached and the fluorescence recovery is measured to generate curves like what is shown in (c). (c) Before photobleaching, the fluorescence is 100%, and the immediate recovery of fluorescence over the first 20 s shows the reassociation of proteins with the bleached structure. (els.net)
  • This interactive tutorial explores variations in photobleaching rates in single, dual, and multiply labeled fluorescence specimens. (fsu.edu)
  • d ) Rate and efficiency of PAmCherry1 photoactivation at different physiological pH values, normalized to the fluorescence intensities at the respective pH (data shown in c ). ( e,f ) Photoactivation ( e ) and photobleaching ( f ) kinetics for the indicated proteins using arc lamp illumination through ×60 oil objective with 390/40 nm ( e ) and 570/30 nm ( f ) filters. (nih.gov)
  • is something called fluorescence recovery after photobleaching called FRAP. (coursera.org)
  • This webinar focuses on light-sheet microscopy, in particular Luxendo's InVi-SPIM, and how this technique is less susceptible to phototoxicity and photobleaching experienced by traditional confocal microscopy techniques. (bruker.com)
  • Alexander Jablonski (1898-1980) - Born in the Ukraine in 1898, Alexander Jablonski is best known as the father of fluorescence spectroscopy. (fsu.edu)
  • Principles of Fluorescence Spectroscopy. (currentprotocols.com)
  • Fluorescence Spectroscopy: An Introduction for Biology and Medicine. (currentprotocols.com)
  • Nanosecond fluorescence spectroscopy of macromolecules. (currentprotocols.com)
  • Dispersion-relation fluorescence spectroscopy of mouse embryonic fibroblast (MEF): a) fluorescence image showing a cell whose actin was labeled with GFP. (phys.org)
  • It provides more information than existing methods, such as fluorescence correlation spectroscopy (FCS), which is widely used for studying molecular transport and diffusion coefficients at a fixed spatial scale. (phys.org)
  • Popescu said the multiplicity of scales the method offers over techniques like fluorescence correlated spectroscopy is key. (phys.org)
  • Multiphoton fluorescence microscopy is a powerful tool combining the techniques of laser scanning microscopy with long wavelength multiphoton fluorescence excitation to capture high-resolution and 3-D images of specimens. (fsu.edu)
  • Because fluorescence consists of photons at a longer wavelength than the excitation radiation, the fluorescence signal can be easily separated from the excitation light using a beamsplitter and/or filters. (imperial.ac.uk)
  • The technique of dual-wavelength ratio fluorescence microscopy provides a powerful tool to measure organellar pH. (omicsonline.org)
  • a) In a primary immunofluorescence assay, an antibody that is directly conjugated to a fluorophore recognises its antigen, and its position is directly detected by fluorescence microscopy. (els.net)
  • Lowest fluorophore concentrations - There is an ongoing drive in fluorescence microscopy to push to lower and lower fluorophore concentrations in order not to perturb the physiology of the living cells being studied. (environmental-expert.com)
  • Three-photon excitation fluorescence axial images are shown of polystyrene beads stained with the fluorophore 2,5-bis(4-biphenyl)oxazole (BBO). (spiedigitallibrary.org)
  • Fluorescence microscopy utilizing multiple low-energy photons to produce the excitation event of the fluorophore. (jove.com)
  • Fluorescence microscopy has enjoyed a renaissance over the last decade, partly driven by advances in light source and detector technologies and partly due to advances in labelling technologies such as fluorescent proteins that can tag specific proteins of interest using genetic engineering. (imperial.ac.uk)
  • The reliance of modern microscopy techniques on photoactivatable fluorescent proteins prompted development of mCherry variants that are initially dark but become red fluorescent after violet-light irradiation. (nih.gov)
  • b ) Maturation kinetics for the indicated proteins at 37 °C. ( c ) Equilibrium pH dependence for the red fluorescence of the indicated proteins. (nih.gov)
  • In this report, we describe a technology for tracking and mapping proteins by multiscale microscopy. (pnas.org)
  • There are various genetically encoded tags, such as fluorescent proteins, developed for fluorescence microscopy (FM). (pnas.org)
  • However, there are almost no genetically encoded tags that enable cellular proteins to be observed by both FM and electron microscopy (EM). (pnas.org)
  • Supercritical angle fluorescence microscopy (SAF) is a technique to detect and characterize fluorescent species (proteins, biomolecules, pharmaceuticals, etc.) and their behaviour close or even adsorbed or linked at surfaces. (wikipedia.org)
  • We have developed and applied a method unifying fluorescence microscopy and mass spectrometry for studying spatial and temporal properties of proteins and protein complexes in yeast cells. (mcponline.org)
  • Fluorescent proteins have become invaluable probes for studying molecular processes in living cells with light microscopy techniques ( 1 - 3 ). (mcponline.org)
  • Super-resolution (SR) fluorescence microscopy encompasses a variety of techniques that can exceed the diffraction limit - the point at which molecular nanostructures can be distinguished from one another. (news-medical.net)
  • The reported performance of diffraction-unlimited fluorescence microscopy opens up a pathway for addressing fundamental problems in the life sciences. (pnas.org)
  • The fluorescence collected during the 15,000 frames is shown in a and represents a diffraction-limited TIRF microscopy image. (nih.gov)
  • Here, we present a compressed sensing approach that allows 3D sub-diffraction nSIM of cultured cells by saturating fluorescence excitation. (nature.com)
  • A fraction of the immobilized polymers is in a clustered or entangled state, which we identify based on diffraction limited fluorescence images. (tue.nl)
  • Image past the so-called diffraction limit of normal fluorescence microscopy. (coursera.org)
  • Using an aspheric lens for excitation of a sample with laser light, fluorescence emitted by the specimen is collected above the critical angle of total internal reflection selectively and directed by a parabolic optics onto a detector. (wikipedia.org)
  • In this interview from SfN 2018, Michelle Gal explains the fluorescence illumination systems for fluorescence microscopy, offered by Excelitas technologies. (news-medical.net)
  • Revenue forecasts from 2018 to 2023 are given for each fluorescence microscopy segment, and regional market with estimated values derived from the manufacturers' total revenues. (bccresearch.com)
  • Revenue forecasts from 2018 to 2023 are given for each fluorescence microscopy segment. (bccresearch.com)
  • The range of available wavelengths for fluorescence excitation has expanded and leads to new combinations when customizing the Colibri.2 for different applications. (nanowerk.com)
  • Confocal microscopy offers the ability to control depth of field, elimination or reduction of background information away from the focal plane, and the capability to collect serial optical sections from thick specimens. (fsu.edu)
  • Researchers have developed a technique that enables visualization of cellular organelles at a resolution that has not previously been achievable in optical microscopy. (news-medical.net)
  • Such fields include scanned beam fluorescence microscopy, scanned beam microlithography, nanofabrication, and optical digital information storage and retrieval. (google.com)
  • Its advantages are based upon attributes not as readily available in other optical microscopy techniques. (fsu.edu)
  • Scientists at Heidelberg Universitys Institute of Physical Chemistry and members of the Cluster of Excellence CellNetworks have devised a new method in which light-dependent processes are replaced by chemical reactions to mark cellular structures for high-resolution optical microscopy. (uni-protokolle.de)
  • In: Masters BR and So PTC (eds) Handbook of Biomedical Nonlinear Optical Microscopy, pp. 191-216. (els.net)
  • We therefore correlate AFM and fluorescence microscopy to couple nanoscale morphological information to far-field optical images. (tue.nl)
  • J. Enderlein, T. Ruckstuhl, S. Seeger: Highly Efficient Optical Detection of Surface-Generated Fluorescence. (wikipedia.org)
  • The appendices provide a quick reference to optical theory, microscopy-related formulas and definitions, and Fourier theory. (spie.org)
  • Its unique feature is that it combines optical techniques -- which are noninvasive, and can be used inside (intact) watery structures -- with high resolution 3D microscopy. (spie.org)
  • The goal of this course was to introduce students with a biology background to some of the fundamental concepts of image formation in confocal fluorescence microscopy and to make them aware of experiment related issues, such as optical aberrations, bleaching, point spread function measurement and digitization. (spie.org)
  • In addition to this, the text was used to compile, over the years, optical formulas and relations that are used in microscopy related from a multitude of sources. (spie.org)
  • In this way the book also serves as a quick reference to optical theory in general and its application to confocal fluorescence microscopy in particular. (spie.org)
  • The focus was on developments in quantitative optical microscopy techniques, which are revolutionizing research in the biological sciences today. (royalsocietypublishing.org)
  • Glossary of Related Terminology with Page References to the Handbook of Optical Filters for Fluorescence Microscopy. (currentprotocols.com)
  • Handbook of Optical Filters for Fluorescence Microscopy. (currentprotocols.com)
  • WITec is a manufacturer of high-resolution optical & scanning probe microscopy solutions for scientific and industrial applications. (spectroscopyeurope.com)
  • Dr. Packer will review his recent work in which he utilized a Spatial Light Modulator (SLM) interfaced to a multiphoton microscopy system in order to simultaneously use optical stimulation and optical recording to study neural networks. (bruker.com)
  • A Shuttle and Find plugin, which allows for correlative light and electron microscopy ( CLEM ) with our Electron Microscopy Unit. (sun.ac.za)
  • Traditionally, biologists have been confined to transmission electron microscopy (TEM) and light microscopy (LM) in order to correlate biochemical and molecular data with morphology. (springer.com)
  • Electron microscopy (EM) provides fine ultrastructural detail but is limited to the study of cellular structures that react with electron-dense stains deposited in fixed specimens. (springer.com)
  • Artifacts in Biological Electron Microscopy (R.F.E. Crang and K.L. Klomparns, eds. (springer.com)
  • Transmission electron microscopy (TEM) is a critical technique for providing information regarding the organization of cellular and magnetite structures in these microorganisms. (osti.gov)
  • There are microscopic techniques that do not depend on fluorescent probes, but have alternative contrast mechanisms, that is, coherent anti‐Stokes Raman microscopy and second‐harmonic generation microscopy. (els.net)
  • The featured resource is provided as a guide and reference tool for visitors who are exploring the large spectrum of specialized topics in fluorescence and laser scanning confocal microscopy. (fsu.edu)
  • The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. (open.ac.uk)
  • The absorption and subsequent re-radiation of light by organic and inorganic specimens is mainly the result of either fluorescence or phosphorescence. (bccresearch.com)
  • Cervical biopsy specimens of colposcopically normal and abnormal tissues obtained from 15 patients were evaluated by confocal fluorescence microscopy. (spie.org)
  • In this presentation, we will focus on the applications of multiphoton microscopy to skin specimens in different physiological states. (spie.org)
  • Unlike other forms of microscopy that utilise reflected or scattered light, this means that very weak fluorescence signals - down to a single photon - can be detected. (imperial.ac.uk)
  • Denk W, Strickler JH and Webb WW (1990) Two‐photon laser scanning fluorescence microscopy. (els.net)
  • We show experiments proving the feasibility of scanning fluorescence microscopy by three-photon excitation. (spiedigitallibrary.org)
  • Two-color two-photon fluorescence laser scanning microscopy. (biomedsearch.com)
  • Fluorescence is induced by two-color two-photon absorption using the fundamental and the second harmonic of a Ti:Sa femtosecond laser. (biomedsearch.com)
  • Jablonski diagrams showing the difference between linear fluorescence (left) and two-photon excitation (right). (azooptics.com)
  • This non-linear fluorescence response has found application in two-photon and multi-photon fluorescence microscopy systems. (azooptics.com)
  • Prior to the development of SR microscopy techniques, conventional wide-field microscopy was unable to distinguish nanostructures that were smaller than 200 nm. (news-medical.net)
  • Conventional fluorescence microscopy uses a lens to focus a beam of light onto a spot. (photonics.com)
  • The super-resolution techniques allow the observation of many biological structures not resolvable in conventional fluorescence microscopy. (hindawi.com)
  • Epi-fluorescence, or incident light fluorescence, has now become the method of choice in many applications and comprises a large part of this tutorial. (fsu.edu)
  • The featured discussion is intended to aid in understanding the basics of light detection and to provide a guide for selecting a suitable detector for specific applications in fluorescence microscopy. (fsu.edu)
  • The Marine Biological Laboratory in Woods Hole, Massachusetts is offering the first (of an anticipated annual) light-sheet fluorescence microscopy conference and workshop. (mbl.edu)
  • The Virtual Conference (May 9-11, 2021) will bring together innovators in microscopy from academic institutes, industry, and federal laboratories to share and brainstorm the latest developments taking place in light-sheet research and its biological applications. (mbl.edu)
  • In light of this, an ideal representation of fluorescence microscopy images would share the features of adaptation and local gain control with the human visual system. (nature.com)
  • The fluorescence image is formed by the action of the two lenses, known as the objective lens, which captures the light from the sample, and the tube lens that forms a real image at the camera. (imperial.ac.uk)
  • a ) Cells expressing TfR-PAmCherry1 were subjected to low-level 405-nm laser photoactivation, and we simultaneously collected 561-nm-laser-light-excited PAmCherry1 single-molecule fluorescence. (nih.gov)
  • Physical limits in high-resolution light microscopy can be overcome with the aid of chemical reactions. (uni-protokolle.de)
  • According to Dr. Herten, it means that microscopy methods like STORM can be simplified to such an extent that additional laser lines and high light intensities become superfluous. (uni-protokolle.de)
  • Brumberg EM (1959) Fluorescence microscopy of biological objects using light from above. (els.net)
  • A camera that measures meter ranges with centimetre precision using the speed of light as a ruler is capable of measuring nanosecond times with tens or hundreds of picoseconds precision, exactly what we do routinely in fluorescence lifetime sensing. (mdpi.com)
  • Light microscopy continues to reveal the microscopic world at an ever increasing resolution. (mpg.de)
  • Examination of the final film surfaces with regular light microscopy and AFM gives additional information about the film formation.These methods have been used for studying the formation of negatively charged latex films. (diva-portal.org)
  • Biological structures span many orders of magnitude in size, but far-field visible light microscopy suffers from limited resolution. (nih.gov)
  • Common bright field microscopy, standard on most nanomechanical test instruments, utilizes broad-spectrum reflected light to image the sample surface and cannot readily distinguish testing regions of interest on complex inhomogeneous materials. (bruker.com)
  • The contents of this book started out as material for a text that I wrote for the Ph.D. course Confocal Light Microscopy: Fundamentals and Biological Applications, which was first given by me in 1996 at the University of Amsterdam. (spie.org)
  • An important factor in fluorescence applications is the use of low light emissions, to reduce the risk of photo bleaching the sample. (baslerweb.com)
  • Even the Basler Microscopy Software has released its 2.0 version: the graphical user interface can be switched to dark skin mode to reduce the light emissions from the display towards the sample. (baslerweb.com)
  • This is the first title on the topic designed specifically to allow students and researchers with little background in physics to understand both microscopy basics and novel light microscopy techniques. (researchandmarkets.com)
  • Light and Video Microscopy. (researchandmarkets.com)
  • Bright-field microscopy relies on the differences in absorption of light due to differences in densities between various parts of the sample, which for our purposes is a cell. (thermofisher.com)
  • Rotating fluorescence light microscopy footage of a cross-section through papillae in tissue from a tongue. (sciencephoto.com)
  • Rotating fluorescence light microscopy footage of skeletal muscle tissue from a tongue. (sciencephoto.com)
  • In comparison to wide-field microscopy, the resulting image has a two-fold improvement in resolution. (news-medical.net)
  • Lumenera's Fluorescence Series cameras are fully integrated into Image-Pro Premier software with a custom Capture Interface, which gives fluorescence microscopy customers complete access to maximum frame rates, live image adjustments, and all the benefits available from INFINITY cameras. (prweb.com)
  • b) An example of a fluorescence image that combines indirect immunofluorescence (a mouse antibody that detects tubulin and a rhodamine‐labelled anti‐mouse secondary that detects the primary antibody), a GFP‐tagged protein and DAPI staining of DNA to label chromosomes (scale bar, 5 µm). (els.net)
  • The tutorial initializes with a pair of identical fluorescence images appearing in the Unbleached Image and Photobleached Image windows. (fsu.edu)
  • Single-particle tracking is powerful in measuring particle motion from video-microscopy image sequences [ 11 , 12 ]. (hindawi.com)
  • left) Fluorescence image of a mouse femur cross-section injected with calcein fluorochrome to monitor tissue age. (bruker.com)
  • Notice the rings marked with calcein fluorochrome cannot be identified using bright-field image microscopy. (bruker.com)
  • The first five chapters cover the main aspects of confocal fluorescence microscopy: image formation, practical limitations, fluorescence, laser operation, and digitization. (spie.org)
  • In microscopy, we refer to the details visible in a magnified image as resolution . (thermofisher.com)
  • An image of the same field of BPAE cells captured using brightfield (left) and fluorescence (right) microscopy. (thermofisher.com)
  • In confocal fluorescence correlation microscopy (FCM) it is important to ensure that the correlation measurement is actually performed at the chosen location of the three-dimensional image of the specimen. (osapublishing.org)
  • We present a confocal FCM design that provides an automatic real-time readout of the location in the confocal microscopic image, which is aligned with the detector of the fluorescence correlation spectrometer. (osapublishing.org)
  • 1989. Fluorescence Microscopy of Living Cells in Culture, parts A and B. Methods Cell Biol. (currentprotocols.com)
  • Fluorescence microscopy is almost as simple to do as bright-field microscopy, and most often it is more specific. (who.int)
  • Because of this, bright-field microscopy is not great when you want to see many details in your cell. (thermofisher.com)
  • The nuclei were stained with Sytox Green (green fluorescence), while the mucus of goblet cells and the filamentous actin in the brush border were stained with Alexa Fluor 350 wheat germ agglutinin (blue fluorescence) and Alexa Fluor 568 phalloidin (red fluorescence), respectively. (fsu.edu)
  • The green fluorescence shows the neurons and the magenta areas are the nuclei of the cells. (nih.gov)
  • The blue fluorescence shows the edges of the cells. (nih.gov)
  • First measurements of living MIN-6 cells reveal differences between the UV fluorescence lifetimes of the nucleus and cytoplasm. (biomedsearch.com)
  • Fluorescence microscopy gives you the advantage of better resolution by making various structures in the cells contrast better with their neighbors, as well as allowing you to collect images in more than one color. (thermofisher.com)
  • about the instrumentation that you need to do fluorescence microscopy in live cells. (coursera.org)
  • Phys.org)-The ability of fluorescence microscopy to study labeled structures like cells has now been empowered to deliver greater spatial and temporal resolutions that were not possible before, thanks to a new method developed by University of Illinois researcher Gabriel Popescu and Ru Wang from his lab. (phys.org)
  • Yet when using fluoresence microscopy only the colonies transfected with the GOI-EGFP-vector show GFP-fluorescence, while none of the EGFP-transfected HEK293 cells seem to exhibit GFP-fluorescence. (protocol-online.org)
  • If that matters: Cells are put on slides using cytospin centrifugation for fluorescence microscopy. (protocol-online.org)
  • Our results indicate that this approach combining fluorescence microscopy and mass spectrometry into a single method provides a unique perspective into molecular mechanisms regulating composition and dynamic properties of the protein complexes in living cells. (mcponline.org)
  • Capture fine details of a variety of cellular and subcellular assays with this powerful and flexible fluorescent microscopy solution. (moleculardevices.com)