Fluorescence: 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.Spectrometry, Fluorescence: Measurement of the intensity and quality of fluorescence.Microscopy, 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.Microscopy: 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, Electron: 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, Electron, Scanning: 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.Microscopy, Confocal: 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.Fluorescence Polarization: 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.In Situ Hybridization, Fluorescence: 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.Fluorescent Dyes: 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.Fluorescence Resonance Energy Transfer: 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.Microscopy, Fluorescence, Multiphoton: 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.Microscopy, Atomic Force: 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.Microscopy, Electron, Transmission: 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, Immunoelectron: 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.Fluorescence Recovery After Photobleaching: 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).Green Fluorescent Proteins: 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.Luminescent Proteins: 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.Cryoelectron Microscopy: 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, Video: 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.Rhodamines: 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.Kinetics: The rate dynamics in chemical or physical systems.Microscopy, Scanning Tunneling: 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, Polarization: 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.Microscopy, Interference: 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.Tryptophan: 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.Energy Transfer: 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.Fluoresceins: 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.Photons: 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)Carbocyanines: Compounds that contain three methine groups. They are frequently used as cationic dyes used for differential staining of biological materials.Staining and Labeling: 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.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Microscopy, Phase-Contrast: 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.Protein Binding: 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.Photobleaching: 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.Optical Imaging: 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.Protein Conformation: 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).Microscopy, Electron, Scanning Transmission: 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.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Fluorometry: 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.Fluorescent Antibody Technique: 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.Molecular Sequence Data: 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.Lasers: 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.Fluorescein-5-isothiocyanate: 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.Microscopy, Scanning Probe: 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).Fluorescein: 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, Cultured: 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.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Hydrogen-Ion Concentration: 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)Circular Dichroism: 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)Diffusion: 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.Temperature: 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.Image Processing, Computer-Assisted: 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.Calcium: 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, Molecular: Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.Molecular Imaging: 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.Anilino Naphthalenesulfonates: 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.Fluorescence Polarization Immunoassay: 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.2-Naphthylamine: A naphthalene derivative with carcinogenic action.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Amino Acid Sequence: 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.Light: That portion of the electromagnetic spectrum in the visible, ultraviolet, and infrared range.Biophysics: The study of PHYSICAL PHENOMENA and PHYSICAL PROCESSES as applied to living things.Rabbits: 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.Liposomes: 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.Flow Cytometry: 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 Fusion Proteins: 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.Chlorophyll: Porphyrin derivatives containing magnesium that act to convert light energy in photosynthetic organisms.Spectrometry, X-Ray Emission: 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.Actins: 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.Biophysical Phenomena: The physical characteristics and processes of biological systems.Cattle: 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.Sensitivity and Specificity: 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)Surface Properties: Characteristics or attributes of the outer boundaries of objects, including molecules.Immunohistochemistry: Histochemical localization of immunoreactive substances using labeled antibodies as reagents.Cytoplasm: 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)DNA: 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).PhotochemistryModels, Biological: 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.Acrylamide: 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.Escherichia coli: 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.Cell Nucleus: 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)Dansyl Compounds: Compounds that contain a 1-dimethylaminonaphthalene-5-sulfonyl group.Pyrenes: A group of condensed ring hydrocarbons.Protein Structure, Tertiary: 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.Lipid Bilayers: Layers of lipid molecules which are two molecules thick. Bilayer systems are frequently studied as models of biological membranes.Microscopy, Acoustic: 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.Coloring Agents: Chemicals and substances that impart color including soluble dyes and insoluble pigments. They are used in INKS; PAINTS; and as INDICATORS AND REAGENTS.Diphenylhexatriene: 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.Membrane Proteins: 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.Spectrophotometry, Ultraviolet: 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)Nanotechnology: The development and use of techniques to study physical phenomena and construct structures in the nanoscale size range or smaller.Optical Phenomena: LIGHT, it's processes and properties, and the characteristics of materials interacting with it.Imaging, Three-Dimensional: 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.Mutation: 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.Naphthalenesulfonates: A class of organic compounds that contains a naphthalene moiety linked to a sulfonic acid salt or ester.Biosensing Techniques: 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.Scattering, Radiation: 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)Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Bacterial Proteins: Proteins found in any species of bacterium.Protein Transport: 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.Phosphatidylcholines: 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.Protein Denaturation: Disruption of the non-covalent bonds and/or disulfide bonds responsible for maintaining the three-dimensional shape and activity of the native protein.Membrane Fluidity: 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.Optics and Photonics: 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.Quantum Dots: 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.Recombinant Proteins: Proteins prepared by recombinant DNA technology.Models, Chemical: Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.Optical Processes: Behavior of LIGHT and its interactions with itself and materials.Particle Size: Relating to the size of solids.Thermodynamics: 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)Nanoparticles: 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.Time-Lapse 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.Indicators and Reagents: 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)Molecular Structure: 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.Pyridinium Compounds4-Chloro-7-nitrobenzofurazan: 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.HeLa Cells: 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.Reproducibility of Results: 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.Aminolevulinic Acid: A compound produced from succinyl-CoA and GLYCINE as an intermediate in heme synthesis. It is used as a PHOTOCHEMOTHERAPY for actinic KERATOSIS.Mathematics: The deductive study of shape, quantity, and dependence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)Indocyanine Green: A tricarbocyanine dye that is used diagnostically in liver function tests and to determine blood volume and cardiac output.Xanthenes: Compounds with three aromatic rings in linear arrangement with an OXYGEN in the center ring.Polymers: Compounds formed by the joining of smaller, usually repeating, units linked by covalent bonds. These compounds often form large macromolecules (e.g., BIOPOLYMERS; PLASTICS).Biological Transport: 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.Image Cytometry: 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.Cell Line, Tumor: A cell line derived from cultured tumor cells.Laurates: Salts and esters of the 12-carbon saturated monocarboxylic acid--lauric acid.Molecular Probes: 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.Adenosine Triphosphate: 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.Protein Structure, Secondary: 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.Nanostructures: 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.Cricetinae: A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.Equipment Design: Methods of creating machines and devices.Cytoskeleton: The network of filaments, tubules, and interconnecting filamentous bridges which give shape, structure, and organization to the cytoplasm.Protein Folding: Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.Molecular Conformation: The characteristic three-dimensional shape of a molecule.Microtubules: 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.Photosensitizing Agents: 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.Gold: 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.Diagnostic Imaging: 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.Histocytochemistry: 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.Guanidine: 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.Endocytosis: Cellular uptake of extracellular materials within membrane-limited vacuoles or microvesicles. ENDOSOMES play a central role in endocytosis.Membrane Lipids: 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, Ultraviolet: Microscopy in which the image is formed by ultraviolet radiation and is displayed and recorded by means of photographic film.Spectrophotometry: The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.Swine: 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).Transfection: 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.Ethidium: 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.Acridine Orange: 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.Rats, Sprague-Dawley: 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.Macromolecular Substances: 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.Molecular Probe Techniques: The use of devices which use detector molecules to detect, investigate, or analyze other molecules, macromolecules, molecular aggregates, or organisms.Membranes, Artificial: 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.Fluorescent Antibody Technique, Indirect: 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)Dextrans: 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: 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.Water: 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)Organic Chemicals: 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.Peptides: 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.Protoporphyrins: 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.Metal Nanoparticles: 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.Spectroscopy, Near-Infrared: 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.Freeze Fracturing: 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.Infrared Rays: 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.Calibration: 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.Mitochondria: 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)Cell Survival: 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.Spectrum Analysis, Raman: Analysis of the intensity of Raman scattering of monochromatic light as a function of frequency of the scattered light.Protein Multimerization: The assembly of the QUATERNARY PROTEIN STRUCTURE of multimeric proteins (MULTIPROTEIN COMPLEXES) from their composite PROTEIN SUBUNITS.Acrylamides: 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.Proteins: 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.Quinolinium CompoundsEquipment Failure Analysis: 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, Monoclonal: Antibodies produced by a single clone of cells.Cryoultramicrotomy: 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.Microspheres: Small uniformly-sized spherical particles, of micrometer dimensions, frequently labeled with radioisotopes or various reagents acting as tags or markers.Polymerase Chain Reaction: 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.Blotting, Western: 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.Phospholipids: 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.Dimerization: The process by which two molecules of the same chemical composition form a condensation product or polymer.Mice, Inbred C57BLLenses: Pieces of glass or other transparent materials used for magnification or increased visual acuity.Cytological Techniques: Methods used to study CELLS.Tomography, Optical: 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.Spectrum Analysis: 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, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Actin Cytoskeleton: Fibers composed of MICROFILAMENT PROTEINS, which are predominately ACTIN. They are the smallest of the cytoskeletal filaments.Micelles: 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.Oxidation-Reduction: 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).Boron Compounds: Inorganic or organic compounds that contain boron as an integral part of the molecule.Color: The visually perceived property of objects created by absorption or reflection of specific wavelengths of light.Microtomy: 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.Chromatography, High Pressure Liquid: Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.Dose-Response Relationship, Drug: The relationship between the dose of an administered drug and the response of the organism to the drug.Europium: 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.Viscosity: 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)Ligands: 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)1,2-Dipalmitoylphosphatidylcholine: Synthetic phospholipid used in liposomes and lipid bilayers to study biological membranes. It is also a major constituent of PULMONARY SURFACTANTS.

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 ...
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 ...
... is a widely applied imaging technique used to examine cells and investigate their internal structures.
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 ...
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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
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…
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 ...
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.
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 ...
​During this seminar, fluorescence microscopy techniques that are available within Shared Research Facilities will be highlighted.
Reflected light fluorescence microscopy is overwhelmingly the current method of choice for widefield investigations with non-coherent light sources, as well as those conducted with laser scanning confocal and multiphoton instruments. Reflected light fluorescence microscopy is overwhelmingly the current method of choice for widefield investigations with non-coherent light sources, as well as those conducted with laser scanning confocal and multiphoton instruments.
Widefield fluorescent image of epithelial cells (nuclei stained with DAPI, yellow; and filamentous actin stained with Alexa Fluor 488 phalloidin, magenta )
In the recent past, a variety of fluorescence microscopy methods emerged that proved to bypass a fundamental limit in light microscopy, the diffraction barrier. Among diverse methods that provide subdiffraction spatial resolution, far-field microscopic techniques are in particular important as they can be operated in complex biological samples such as cells or tissue. Valuable new insights into biomolecular structure, organization and even dynamic processes in living cells have been gained with these novel microscopic techniques. In the present review, the most important concepts of far-field microscopy with subdiffraction resolution are introduced. The underlying physical concepts are discussed, and practical considerations for the application of these methods are made. (C) 2010 Elsevier B.V. All rights reserved ...
This page shows application examples of KEYENCE BZ-X Series biological fluorescence microscopes. The BZ-X Fluorescence Microscope supports bright field, phase contrast, oblique illumination, and fluorescence observation all with this single unit and without the need for a darkroom.
This page shows application examples of KEYENCE BZ Series biological fluorescence microscopes. The BZ-X700 Fluorescence Microscope supports bright field, phase contrast, oblique illumination, and fluorescence observation all with this single unit and without the need for a darkroom.
We revisited the adaptive response to DNA methylation damage that was discovered in the bacterium Escherichia coli back in 1977, but using modern super-resolution fluorescence microscopy and single-cell microfluidic imaging. To our surprise, we found that the Ada protein required to sense DNA methylation damage was present at extremely low quantities in cells before damage. Cells had on average only a single copy of Ada, and a significant fraction of the cells had none at all. How many copies each cell contained was purely due to chance - or bad luck for those cells that had none. It turned out that the cells with no copies did not sense the presence of DNA damage. They were thus unable to activate the adaptive response that would allow them to repair the damage ...
The spliceosome found in human cells is made up of many different subunits, which must be assembled onto the mRNA precursor in a series of carefully choreographed steps. The binding specificity of individual subunits is crucial for both spliceosome assembly and function. "The assembly factor we have studied, called the U2 Auxiliary Factor or U2AF for short, is critical for the correct recognition of the splicing sites at one end of the introns," says Lena Voithenberg, first author of the new paper. U2AF itself is made up of two different subunits. In its free form, the larger of the two is a highly dynamic protein, as Voithenberg and her colleagues demonstrated by means of single-molecule fluorescence microscopy. Experiments using nuclear magnetic resonance (NMR) spectroscopy carried out in parallel at the Bavarian Center for NMR (run jointly by the Helmholtz Zentrum München and the TUM) provided further information relating to the structure and conformational dynamics of U2AF. ...
DPhil Candidate Marko Sustarsic is a DPhil student in Structural Biology and is based at the Department of Physics at the University of Oxford. He is working on an interdisciplinary project that employs single-molecule fluorescence microscopy to explore the structure and dynamics of DNA-binding proteins in bacteria. Contact email
Using fluorescence microscopy with single molecule sensitivity it is now possible to follow the movement of individual fluorophore tagged molecules such as proteins and lipids in the cell membrane with nanometer precision. These experiments are important as they allow many key biological processes on the cell membrane and in the cell, such as transcription, translation and DNA replication, to be studied at new levels of detail. Computerized microscopes generate sequences of images (in the order of tens to hundreds) of the molecules diffusing and one of the challenges is to track these molecules to obtain reliable statistics such as speed distributions, diffusion patterns, intracellular positioning, etc. The data set is challenging because the molecules are tagged with a single or small number of fluorophores, which makes it difficult to distinguish them from the background, the fluorophore bleaches irreversibly over time, the number of tagged molecules are unknown and there is occasional loss of ...
Specimens labelled with multiple fluorophores illuminating a variety of cellular targets, facilitated by developments in probes, data acquisition and analysis technologies, have extended the capabilities of anatomical investigations. However, the field has largely neglected development and consensus in optimal techniques of specimen preparation. The optical properties of the specimen largely determines study outcome in epifluorescence microscopy. Thus, we discuss tissue preparation protocols and their application in multiple anatomical investigations. The processing of tissues optimised for multiple studies are detailed. The conservation of cyto-architecture and antigenicity, reduced tissue autofluorescence, increased permeabilisation, and applications for thick tissue sections are examined. Multiple analytical assays of the same tissue sections for the cross correlation of multiple data sets are presented. The application of multiple labelling immunofluorescence, fluorescent in situ ...
We recently finished our Ask the Expert discussion on Improving Live Cell Fluorescence Imaging. This week we had several interesting questions focused on combating the negative effects of imaging on cell health and viability including looking at media options as a possible solution.
Immunofluorescent Staining:. Nerves, capillaries, and basement membranes are localized by immunohistochemistry in our laboratory. Selected antibodies bind with high specificity to relevant proteins in the sectioned skin biopsy. Fluorescently-labeled secondary antibodies are then applied to the tissue sections and attach to the previously applied antibodies, forming labeled antibody complexes that can be observed with a fluorescent microscope. By using different antibodies that have been localized with distinct fluorescent dyes, multiple structures can be examined in the same section.. After the slides are stained, we view them with a fluorescent microscope. It uses a highly sensitive camera that collects images that can be recorded for image analysis and quatification of nerve fibers.. Confocal Microscopy:. A lot of our work is based on confocal microscopy where we collect a series of images (16 to 60) through a thick section of tissue -- much like a CT scan. Each image focuses on a thin plane ...
The Olympus BioScapes International Digital Imaging Competition is an international contest to find and honor the worlds most astonishing microscopy images of life science subjects.
The Microscopy Core Facility supports both research and teaching needs. Resources include a scanning electron microscope (SEM), a transmission electron microscope (TEM), a laser scanning confocal microscope, and a direct epifluorescence microscope equipped with a digital CCD camera to allow for FRAP, FRET and high-speed multi-channel time-lapse analyses. This facility is also staffed by a full-time research technician to provide training, oversight, and technical support.. To use shared microscope facilities, contact Dr. Abigail Reft, Biology Department Microscopist. Read more about available Microscopes and Sample Preparation. ...
This patent search tool allows you not only to search the PCT database of about 2 million International Applications but also the worldwide patent collections. This search facility features: flexible search syntax; automatic word stemming and relevance ranking; as well as graphical results.
Yakimovich, A; Gumpert, H; Burckhardt, C J; Lutschg, V A; Jurgeit, A; Sbalzarini, I F; Greber, U F (2012). Cell-free transmission of human adenovirus by passive mass transfer in cell culture simulated in a computer model. Journal of Virology, 86(18):10123-10137.. Cardinale, J; Paul, G; Sbalzarini, I F (2012). Discrete region competition for unknown numbers of connected regions. IEEE transactions on image processing : a publication of the IEEE Signal Processing Society, 21(8):3531-3545.. Helmuth, J A; Burckhardt, C J; Greber, U F; Sbalzarini, I F (2009). Shape reconstruction of subcellular structures from live cell fluorescence microscopy images. Journal of Structural Biology, 167(1):1-10.. Helmuth, J A; Burckhardt, C J; Koumoutsakos, P; Greber, U F; Sbalzarini, I F (2007). A novel supervised trajectory segmentation algorithm identifies distinct types of human adenovirus motion in host cells. Journal of Structural Biology, 159(3):347-358.. ...
(a) Representative in vivo fluorescence images of MGC803-tumour-bearing mouse after iv-injected with FA-AlexaFluor647-labeled pRNA nanoparticle. The tumor areas
Display event corresponding to Figure 4A during pH oscillations, recorded at 10 Hz with a wide field epifluorescence microscope. Opening (sudden increase in fluorescence which oscillates with pH, green arrow) and closing (the fluorescence of the structure stops oscillating, orange arrow) are clearly visible. Movie is slowed down 2 times, scale bar is 1 µm. ...
Imaging apparatus and method which uses change of polarization state of a light beam passed through a total internal reflection structure by a single reflection at a TIR surface in which a specimen is placed in the evanescent field associated with the total internal reflection of the light beam, the specimen being the subject of biological, chemical or genetic investigation.
X-Cite FIRE light source for fluorescence microscopy offers a rich, broad spectrum output from 360-750nm, for exciting an extended range of fluorophores wi...
Optical Fluorescence Microscopy. From the Spectral to the Nano Dimension. Editors: Diaspro, Alberto (Ed.) Free Preview. Offers a modern approach to.
Optical Fluorescence Microscopy von Alberto Diaspro und Buchbewertungen gibt es auf ReadRate.com. Bücher können hier direkt online erworben werden.
Molecular Probes® brand fluorescent products are recognized the world over as key components for quality fluorescent images, both in our publications and in yours. Many reagents and imaging refinements go into the production of a biologically relevant and informative image. After the use of functional probes, labels, and counterstains, additional tools are often needed to achieve a stunning cellular image.
usually you use the one, with which you have already experiences. And of course also something what you have equipment for. E.g. if you do not have fluorescent microscope, you probably wont want to use GFP ...
EWF10x focusing eyepieces with eyeguards, 22mm field of view Siedentopf trinocular viewing head with photo tube; inclined 30º, rotatable 360°
One of the newest advances in microscopy technology providing super-resolution are hyperlenses. They make use of so-called evanescent waves that are lost in conventional lenses.
Formed in 1993 by owner and MD Ian Corless, Image Solutions has carved a niche for itself as a consultancy capable of putting together the type of imaging packages demanded by cutting-edge researchers: particularly those who work with live ...
In meeting the goals and objectives of FMIC, the center continues to: -Service and maintain state-of-the-art equipment for unlimited access by intramural scient...
Hoyer P, de Medeiros G, Balázs B, Norlin N, Besir C, Hanne J, Kräusslich HG, Engelhardt J, Sahl SJ, Hell SW, Hufnagel L. (2016). Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT.. Proc. Natl. Acad. Sci. U.S.A. 113(13):3442-3446. doi: 10.1073/pnas. ...
Aqueous mounting medium for use in fluorescence microscopy. Enhances FITC intensity and reduces fluorescence fading. Ready-to-use. Reagent: 6 mlProcedure: FOR USE IN FLUORESCENCE MICROSCOP…
Tools for 4D nucleome imaging. Quantitative analysis of the 3D nuclear landscape recorded with super-resolved fluorescence microscopy.. ...
GeneCone chambers, HybriWell sealing chambers, ONCYTE Film-WellT for cell based microarrays, HybriSlip hybridization cover, MultiSlip cell culture coverslip inserts, Secureslip cell Culture coverslips, CultureWell chambered coverglass plate inserts
Define wide-field fluorescence microscopy: Wide-field fluorescence microscopy uses either naturally occurring structures or staining with fluorescent ...
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 ...
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, ...
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 ...
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.. ...
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) ...
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 ...
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.
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?
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.
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
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 ...
The BD Accuri C6 flow cytometer can be used for a variety of experimental setups, such as 4-parameter cell phenotyping, apoptosis detection (which includes DNA fragmentation and analysis of mitochondrial membrane potential), bacterial and environmental sample analysis, plant ploidy, and cell cycle analysis in mammalian cells.. The system is easy to use and ideal for most novice flow cytometry users, as they can collect and analyse data in a short amount of time. Software options and instrument controls are clearly visible from the softwares tabbed interface which enables access to the collection, analysis, and statistics functions.. The BD Accuri C6 software allows for live gating, event colouring, export of publication-quality graphics, and batch analysis, for review or modification of multiple samples for the automatic creation of Microsoft PowerPoint® and Excel files.. This will allow more freedom to users as minimal assistance is required for running the instrument.. ...
Fluorescent microscopy is a technique where a lower wavelength of light is applied to a sample, but only the light emitted at a higher wavelength is detected. Fluorescent microscopy allows for the sensitive visualization of fluorescent molecules against a dark background. Let Protein Mods label your primary antibody, so you can avoid wasting time with secondary antibodies. In addition to services, Protein Mods offers a number of proteins modified with fluorescein, tetramethylrhodamine, and CY5.5 for your fluorescent microscopy needs. Additional fluorophore options are available upon request. Anti-rabbit IgG Anti-mouse IgG Avidin Protein A Negative controls available in our catalog: Rabbit IgG Mouse IgG Canine IgG Rat IgG BSA
Directional migration requires the coordination of cytoskeletal changes essential for cell polarization and adhesion turnover. Extracellular signals that alter tyrosine phosphorylation drive directional migration by inducing reorganization of the actin cytoskeleton. It is recognized that Nck is an important link between tyrosine phosphorylation and actin dynamics, however, the role of Nck in cytoskeletal remodeling during directional migration and the underlying molecular mechanisms remain largely undetermined. In this study, a combination of molecular genetics and quantitative live cell microscopy was used to show that Nck is essential in the establishment of front-back polarity and directional migration of endothelial cells. Time-lapse differential interference contrast and total internal reflection fluorescence microscopy showed that Nck couples the formation of polarized membrane protrusions with their stabilization through the assembly and maturation of cell-substratum adhesions. ...
Purpose: : Drusen play a key role in AMD, but very little is known about composition and the autofluorescent (AF) material sometimes found within drusen. Here we present a detailed analysis of drusen containing autofluorescent material using Structured Illumination Microscopy (SIM), which provides a lateral resolution twice as high as conventional fluorescence microscopy. Methods: : Ten histological RPE sections obtained from ten human donor eyes (range 57 to 95 years) were examined by SIM using laser light of different wavelengths (488, 568, and 647 nm). Drusen were studied regarding size and shape. AF material within drusen was analyzed in terms of size, shape, AF behaviour and distribution across drusen. Results: : A total of 400 drusen were found, of which 101 contained AF material. 90 % of these drusen were smaller than 63 µm (mean: 35.63 µm ± 0.25 µm). AF material within drusen were identified as lipofuscin (LF, n=183) and melanolipofuscin (MLF, n=36) granules. Up to 11 LF and/or 3 MLF ...
Kurtz, R.: Bright Solutions to Get Sharp Images: Confocal and Two-Photon Fluorescence Microscopy and the Pros and Cons of New Multifocal Approaches. In: Mendez-Vilas, A. and Diaz, J. (eds.) Modern Research and Educational Topics in Microscopy. Microscopy Series No. 3 Vol. 1. p. 154-163. Formatex (2007 ...
TY - JOUR. T1 - UNC-45A is a novel microtubule-associated protein and regulator of paclitaxel sensitivity in ovarian cancer cells. AU - Mooneyham, Ashley. AU - Iizuka, Yoshie. AU - Yang, Qing. AU - Coombes, Courtney. AU - McClellan, Mark. AU - Shridhar, Vijayalakshmi. AU - Emmings, Edith. AU - Shetty, Mihir. AU - Chen, Liqiang. AU - Ai, Teng. AU - Meints, Joyce. AU - Lee, Michael K.. AU - Gardner, Melissa. AU - Bazzaro, Martina. PY - 2019/2/1. Y1 - 2019/2/1. N2 - UNC-45A, a highly conserved member of the UCS (UNC45A/CRO1/SHE4P) protein family of cochaperones, plays an important role in regulating cytoskeletal-associated functions in invertebrates and mammalian cells, including cytokinesis, exocytosis, cell motility, and neuronal development. Here, for the first time, UNC-45A is demonstrated to function as a mitotic spindle-associated protein that destabilizes microtubules (MT) activity. Using in vitro biophysical reconstitution and total internal reflection fluorescence microscopy analysis, we ...
TY - JOUR. T1 - Nanosecond fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy to localize the protein interactions in a single living cell. AU - Elangovan, M.. AU - Day, R. N.. AU - Periasamy, A.. PY - 2002/2/18. Y1 - 2002/2/18. N2 - Visualizing and quantifying protein-protein interactions is a recent trend in biomedical imaging. The current advances in fluorescence microscopy, coupled with the development of new fluorescent probes such as green fluorescent proteins, allow fluorescence resonance energy transfer (FRET) to be used to study protein interactions in living specimens. Intensity-based FRET microscopy is limited by spectral bleed-through and fluorophore concentration. Fluorescence lifetime imaging (FLIM) microscopy and lifetime measurements are independent of change in fluorophore concentration or excitation intensity, and the combination of FRET and FLIM provides high spatial (nanometre) and temporal (nanoseconds) resolution. Because only the donor ...
TY - JOUR. T1 - Colocalization of connexin 43 and connexin 45 but absence of connexin 40 in granulosa cell gap junctions of rat ovary. AU - Okuma, A.. AU - Kuraoka, A.. AU - Iida, H.. AU - Inai, T.. AU - Wasano, K.. AU - Shibata, Y.. PY - 1996/7. Y1 - 1996/7. N2 - The expression and localization of gap junction family proteins (connexins) were examined in nonstimulated and gonadotrophin-stimulated ovarian follicles of immature rats. Immunoblot and RNA blot analysis showed the presence of connexin (Cx) 43, Cx40 and Cx45 in ovarian tissue. Of these connexin proteins, Cx43 and Cx45 were identified by immunofluorescent microscopy between granulosa cells in characteristic expression patterns related to follicular developmental stages, while Cx40 was not expressed in granulosa cells but was detected in blood vessels in ovarian stroma. In some plaques of gap junction between granulosa cells, Cx45 was found to be colocalized with Cx43. In immunofluorescent microscopy, the expression of Cx43 was ...
Tissue sampling is a problematic issue for inflammatory breast carcinoma, and immediate evaluation following core needle biopsy is needed to evaluate specimen adequacy. We sought to determine if confocal fluorescence microscopy provides sufficient resolution to evaluate specimen adequacy by comparing invasive tumor cellularity estimated from standard histologic images to invasive tumor cellularity estimated from confocal images of breast core needle biopsy specimens. Grayscale confocal fluorescence images of breast core needle biopsy specimens were acquired following proflavine application. A breast-dedicated pathologist evaluated invasive tumor cellularity in histologic images with hematoxylin and eosin staining and in grayscale and false-colored confocal images of cores. Agreement between cellularity estimates was quantified using a kappa coefficient. 23 cores from 23 patients with suspected inflammatory breast carcinoma were imaged. Confocal images were acquired in an average of less than 2 ...
TY - JOUR. T1 - Investigating protein-protein interactions in living cells using fluorescence lifetime imaging microscopy. AU - Sun, Yuansheng. AU - Day, Richard. AU - Periasamy, Ammasi. PY - 2011/9. Y1 - 2011/9. N2 - Fluorescence lifetime imaging microscopy (FLIM) is now routinely used for dynamic measurements of signaling events inside living cells, including detection of protein-protein interactions. An understanding of the basic physics of fluorescence lifetime measurements is required to use this technique. In this protocol, we describe both the time-correlated single photon counting and the frequency-domain methods for FLIM data acquisition and analysis. We describe calibration of both FLIM systems, and demonstrate how they are used to measure the quenched donor fluorescence lifetime that results from FÃ ¶rster resonance energy transfer (FRET). We then show how the FLIM-FRET methods are used to detect the dimerization of the transcription factor CCAAT/enhancer binding protein-Î ± in ...
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 ...
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 ...
Yuste R (December 2005). "Fluorescence microscopy today". Nature Methods. 2 (12): 902-4. doi:10.1038/nmeth1205-902. PMID ... cryo-electron microscopy روش دیگری است برای مطالعه ساختار پروتئین که معمولاً در دمای نزدیگ به نیتروژن مایع انجام می‌شود. ... 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 ...
... super-resolved fluorescence microscopy). Referensi[sunting , sunting sumber]. *^ "Eighty-Ninth Annual Commencement - California ...
"Phototoxicity in live fluorescence microscopy, and how to avoid it". BioEssays. 39 (8): 1700003. doi:10.1002/bies.201700003. ... Phototoxicity in light microscopy[edit]. When performing microscopy on live samples, one needs to be aware that too high light ... "Assessing phototoxicity in live fluorescence imaging". Nature Methods. 14 (7): 657-661. doi:10.1038/nmeth.4344. hdl:21.11116/ ...
Often used in fluorescence microscopy for DNA staining, Hoechst stains appear yellow when dissolved in aqueous solutions and ... Electron microscopy[edit]. As in light microscopy, stains can be used to enhance contrast in transmission electron microscopy. ... Rhodamine is a protein specific fluorescent stain commonly used in fluorescence microscopy. ... Wikimedia Commons has media related to Microscopy staining methods.. *[1] The Biological Stain commission is an independent non ...
Raman microscopy, and in particular confocal microscopy, has very high spatial resolution. For example, the lateral and depth ... At wavelengths of 260 nm, there is effectively no fluorescence interference and the UV signal is inherently strong.[28][64][65] ... Raman microscopy for biological and medical specimens generally uses near-infrared (NIR) lasers (785 nm diodes and 1064 nm Nd: ... Raman microscopy of inorganic specimens, such as rocks and ceramics and polymers, can use a broader range of excitation ...
Cherukuri, Paul; Bachilo, Sergei M.; Litovsky, Silvio H.; Weisman, R. Bruce (2004). "Near-Infrared Fluorescence Microscopy of ... Nanotube fluorescence is under investigation for biomedical imaging and sensors.[146][147][148] ... Carbon nanotube photoluminescence (fluorescence) can be used to observe semiconducting single-walled carbon nanotube species. ...
The suspension is analysed by fluorescence microscopy, which automatically counts the events. Simultaneous event galleries are ... fluorescence in situ hybridization (FISH) DNA and RNA assays, and all other relevant molecular techniques using DNA and RNA. ... including the fluorescence intensity of the four fluorescent markers and 86 different morphological parameters. Epic can also ...
Electrophoresis at high pH results in structures resembling comets, observed by fluorescence microscopy; the intensity of the ... The image analysis measures the overall intensity of the fluorescence for the whole nucleoid and the fluorescence of the ... This is followed by visual analysis with staining of DNA and calculating fluorescence to determine the extent of DNA damage. ...
ISBN 0-470-05818-8. Rost, F. W. D. (1992). Fluorescence Microscopy. Cambridge University Press. p. 22. ISBN 0-521-23641-X. ... Stokes fluorescence is the emission of a longer-wavelength photon (lower frequency or energy) by a molecule that has absorbed a ... 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. Practical ...
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 ... Yuste R (Dec 2005). "Fluorescence microscopy today". Nature Methods. 2 (12): 902-4. doi:10.1038/nmeth1205-902. PMID 16299474. ... 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. doi:10.1038/nmeth1205-902. PMID 16299474. ... 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 ... ISBN 0-521-65873-X. Mayhew TM, Lucocq JM (2008). "Developments in cell biology for quantitative immunoelectron microscopy based ...
Fluorescence near-field scanning optical microscopy (NSOM) utilizing an integrated QD-LED has been demonstrated.[150] ... Hoshino, K.; Gopal, A.; Glaz, M. S.; Vanden Bout, D. A.; Zhang, X. (2012). "Nanoscale fluorescence imaging with quantum dot ... The tunability of emission wavelengths and narrow bandwidth is also beneficial as excitation sources for fluorescence imaging. ... cerium doped phosphor coating on the emitter absorbs some of the blue emission and produces yellow light through fluorescence. ...
... an immunohistochemistry-laser scanning confocal fluorescence microscopy study". The Journal of Neuroscience. 14 (11 pt. 2): ... Carlén, B.; Englund, E. (August 2001). "Diagnostic value of electron microscopy in a case of juvenile neuronal ceroid ...
... 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 ... Leung, Bonnie O.; Chou, Keng C. (2011-09-01). "Review of Super-Resolution Fluorescence Microscopy for Biology". Applied ...
Rusk, N. (2009). "The fluorescence microscope". Milstones 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 ...
... 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 ...
Juan Carlos Stockert, Alfonso Blázquez-Castro (2017). "Chapter 3 Dyes and Fluorochromes". Fluorescence Microscopy in Life ... fluorescence)) or to relay their fluorescence at even longer wavelength (see article FRET) See more on fluorescence principle. ... Tsien RY; Waggoner A (1995). "Fluorophores for confocal microscopy". In Pawley JB. Handbook of biological confocal microscopy. ... Fluorescence Microscopy in Life Sciences. Bentham Science Publishers. pp. 96-134. ISBN 978-1-68108-519-7. Retrieved 24 December ...
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." Pawley JB (2006). ... 2013). "Bridging the gap between qualitative and quantitative colocalization results in fluorescence microscopy studies". Sci ...
Absolute fluorescence sensitivity is generally lower in confocal microscopy because out-of-focus signals are rejected by the ... "Sorting Out Fluorescence Activated Cell Sorting". Retrieved 2017-11-09.. *^ Julius MH, Masuda T, Herzenberg LA (1972). " ... Java Fluorescence Spectrum Viewer (Becton, Dickinson and Company). *The History of the Cell Sorter Interviews from the ... The current record for a commercial instrument is ten lasers[6] and 30 fluorescence detectors.[7] Increasing the number of ...
"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. ...
Hepler, P. K.; B. E. S. Gunning (1998). "Confocal fluorescence microscopy of plant cells". Protoplasma. 201 (3): 121-157. doi: ... 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 ... CS1 maint: Multiple names: authors list (link) "Poems and Quotations About the MicroWorld". Microscopy Society of America. ...
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 ...
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: 73-76. Bibcode:1990Sci ... 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 ...
NanoSIMS combined with fluorescence microscopy can be used as a tool for subcellular imaging of isotopically labeled platinum- ... "NanoSIMS combined with fluorescence microscopy as a tool for subcellular imaging of isotopically labeled platinum-based ... NanoSIMS can be combined with transmission electron microscopy (TEM) when using microtome or FIB sections. This combination ...
... 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 ...
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 ...
... 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 ...
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 ...
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 ...
... Glenn White gwhite at ashland.edu Fri Oct 23 10:56:29 EST 1998 *Previous message: senior ...
... 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 ...
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 ...
Fluorescence lifetime imaging (FLIM) can report on photophysical events that are difficult or impossible to observe by ... the fluorescence emission can be characterised not only by intensity and position, but also by lifetime, polarization and ... fluorescence intensity imagi In honour of Hiroshi Masuhara ... Time-resolved fluorescence microscopy Klaus Suhling,*a Paul M. ... fluorescence microscopy. , the fluorescence emission can be characterised not only by intensity and position, but also by ...
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 ...
... - Leica, Olympus - published on openPR.com ... Fluorescence Microscopy Market Worth 530 Million USD by 2023 Globally Fluorescence microscopy is an optical microscope that ... Fluorescence Lifetime Imaging Microscopy production volume, data pertaining to demand and Fluorescence Lifetime Imaging ... You can edit or delete your press release Global Fluorescence Lifetime Imaging Microscopy Market 2018 - Leica, Olympus here. ...
combined total internal reflection fluorescence microscopy with oblique illumination fluorescence microscopy to observe the ... Motion Analysis of Live Objects by Super-Resolution Fluorescence Microscopy. Chunyan Yao,1,2 Jianwei Zhang,2 Guang Wu,3,4 and ... With fluorescence microscopy, it is possible to isolate individual steps along the gene-delivery pathway to characterize the ... B. Huang, M. Bates, and X. Zhuang, "Super-resolution fluorescence microscopy," Annual Review of Biochemistry, vol. 78, pp. 993- ...
  • In addition, important biological applications of FRET microscopy, such as protein interaction studies, Ca 2+ signaling, nucleic acid studies, characterization of gene expression, and real-time PCR assays, are discussed. (rupress.org)
  • A discussion about the successes of other FRET microscopy techniques is also included. (rupress.org)
  • FRET microscopy relies on the ability to capture fluorescent signals from the interactions of labeled molecules in single living or fixed cells. (rupress.org)
  • 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)
  • 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)
  • 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)
  • 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)
  • 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)
  • 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)
  • Lumenera's INFINITY Fluorescence Series cameras have a new sleek black enclosure, and feature Sony CCD sensors to offer high dynamic range, high sensitivity, and optional thermoelectric cooling, resulting in excellent quality images and fast preview speeds. (prweb.com)
  • Recent improvements in methods of single-particle fluorescence tracking have permitted detailed studies of molecular motion on the nanometer scale due to its high selectivity, sensitivity, simplicity, and fastness [ 8 ]. (hindawi.com)
  • The high microscopy sensitivity resolves the signal of a single fluorescence-labeled bimolecular within a living cell. (hindawi.com)
  • Fluorescence is one of the most commonly used physical phenomena in biological and analytical microscopy, mainly because of its high sensitivity and high specificity. (leica-microsystems.com)
  • Based on its increased sensitivity and reduced reading time compared with conventional Ziehl-Neelsen (ZN) light microscopy, the World Health Organization (WHO) recommended the introduction of light-emitting diode fluorescence microscopy (LED-FM) as an alternative to ZN microscopy in both high- and low-volume laboratories [ 3 ]. (ersjournals.com)
  • LED fluorescence microscopy has demonstrated increased sensitivity and. (bio-medicine.org)
  • LED fluorescence microscopy has demonstrated increased sensitivity and speed in numerous research and clinical applications, including the detection of infectious diseases such as tuberculosis and malaria. (bio-medicine.org)
  • Fluorescence microscopy is an excellent method of studying material that can be made to fluoresce, either in its natural form (termed primary or autofluorescence ) or when treated with chemicals capable of fluorescing (known as secondary fluorescence ). (fsu.edu)
  • 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)
  • 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)
  • In this presentation, we will focus on the applications of multiphoton microscopy to skin specimens in different physiological states. (spie.org)
  • The basic idea was to find a silicon-based detector capable of demodulating fluorescence signals on a bi-dimensional array of pixels. (mdpi.com)
  • The review by Petrášek & Schwille (2009) explores the origins of fluctuations in fluorescence signals. (royalsocietypublishing.org)
  • Fluorescence signals are usually indirect measures of the interactions of actual interest. (royalsocietypublishing.org)
  • This paper investigates the expansion of phase analysis to fluorescence self-interference signals to provide functional information about a sample. (osapublishing.org)
  • Combined with genetics and molecular biology techniques fluorescence microscopy provides an efficient tool for observing molecular phenotypes useful for dissecting the pathways of cell cycle progression and cell response to internal and external signals ( 7 ). (mcponline.org)
  • Theoretical sessions will provide fundamentals of microscopy to aid the hands-on-training sessions. (icgeb.org)
  • I would also like to express my appreciation to the department of Molecular Cytology of the Swammerdam Institute for Life Sciences at the University of Amsterdam for giving me the opportunity to teach the confocal microscopy course to Ph.D. students. (spie.org)
  • Increasing the number of measurement parameters from fluorescence experiments maximizes one's ability to interpret the underlying molecular processes. (royalsocietypublishing.org)
  • The Leica M205 FA stereomicroscope is suitable for demanding fluorescence applications in developmental, molecular and cellular biology. (labonline.com.au)