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.
The study of the structure, growth, activities, and functions of NEURONS and the NERVOUS SYSTEM.
Facilities equipped to carry out investigative procedures.
Liquids transforming into solids by the removal of heat.
High-molecular-weight insoluble polymers that contain functional cationic groups capable of undergoing exchange reactions with anions.
Inland bodies of still or slowly moving FRESH WATER or salt water, larger than a pond, and supplied by RIVERS and streams.
Preservation of cells, tissues, organs, or embryos by freezing. In histological preparations, cryopreservation or cryofixation is used to maintain the existing form, structure, and chemical composition of all the constituent elements of the specimens.
A highly contagious herpesvirus infection affecting the central nervous system of swine, cattle, dogs, cats, rats, and other animals.
A species of VARICELLOVIRUS producing a respiratory infection (PSEUDORABIES) in swine, its natural host. It also produces an usually fatal ENCEPHALOMYELITIS in cattle, sheep, dogs, cats, foxes, and mink.
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.
Societies whose membership is limited to physicians.
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.
Vaccines or candidate vaccines used to prevent PSEUDORABIES (Aujeszky's disease), a herpesvirus of swine and other animals.
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.
Electron microscopy in which the ELECTRONS or their reaction products that pass down through the specimen are imaged below the plane of the specimen.
Rare congenital metabolism disorders of the urea cycle. The disorders are due to mutations that result in complete (neonatal onset) or partial (childhood or adult onset) inactivity of an enzyme, involved in the urea cycle. Neonatal onset results in clinical features that include irritability, vomiting, lethargy, seizures, NEONATAL HYPOTONIA; RESPIRATORY ALKALOSIS; HYPERAMMONEMIA; coma, and death. Survivors of the neonatal onset and childhood/adult onset disorders share common risks for ENCEPHALOPATHIES, METABOLIC, INBORN; and RESPIRATORY ALKALOSIS due to HYPERAMMONEMIA.
Stable elementary particles having the smallest known negative charge, present in all elements; also called negatrons. Positively charged electrons are called positrons. The numbers, energies and arrangement of electrons around atomic nuclei determine the chemical identities of elements. Beams of electrons are called CATHODE RAYS.
Research that involves the application of the natural sciences, especially biology and physiology, to medicine.
The main structural coat protein of COATED VESICLES which play a key role in the intracellular transport between membranous organelles. Each molecule of clathrin consists of three light chains (CLATHRIN LIGHT CHAINS) and three heavy chains (CLATHRIN HEAVY CHAINS) that form a structure called a triskelion. Clathrin also interacts with cytoskeletal proteins.
A species of FLAVIVIRUS, one of the Japanese encephalitis virus group (ENCEPHALITIS VIRUSES, JAPANESE). It can infect birds and mammals. In humans, it is seen most frequently in Africa, Asia, and Europe presenting as a silent infection or undifferentiated fever (WEST NILE FEVER). The virus appeared in North America for the first time in 1999. It is transmitted mainly by CULEX spp mosquitoes which feed primarily on birds, but it can also be carried by the Asian Tiger mosquito, AEDES albopictus, which feeds mainly on mammals.
Cellular uptake of extracellular materials within membrane-limited vacuoles or microvesicles. ENDOSOMES play a central role in endocytosis.
A mosquito-borne viral illness caused by the WEST NILE VIRUS, a FLAVIVIRUS and endemic to regions of Africa, Asia, and Europe. Common clinical features include HEADACHE; FEVER; maculopapular rash; gastrointestinal symptoms; and lymphadenopathy. MENINGITIS; ENCEPHALITIS; and MYELITIS may also occur. The disease may occasionally be fatal or leave survivors with residual neurologic deficits. (From Joynt, Clinical Neurology, 1996, Ch26, p13; Lancet 1998 Sep 5;352(9130):767-71)
Vesicles formed when cell-membrane coated pits (COATED PITS, CELL-MEMBRANE) invaginate and pinch off. The outer surface of these vesicles is covered with a lattice-like network of the protein CLATHRIN. Shortly after formation, however, the clathrin coat is removed and the vesicles are referred to as ENDOSOMES.
Proteins, usually glycoproteins, found in the viral envelopes of a variety of viruses. They promote cell membrane fusion and thereby may function in the uptake of the virus by cells.
The entering of cells by viruses following VIRUS ATTACHMENT. This is achieved by ENDOCYTOSIS, by direct MEMBRANE FUSION of the viral membrane with the CELL MEMBRANE, or by translocation of the whole virus across the cell membrane.
The span of viability of a tissue or an organ.
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
The ten-layered nervous tissue membrane of the eye. It is continuous with the OPTIC NERVE and receives images of external objects and transmits visual impulses to the brain. Its outer surface is in contact with the CHOROID and the inner surface with the VITREOUS BODY. The outer-most layer is pigmented, whereas the inner nine layers are transparent.
INTERNEURONS of the vertebrate RETINA. They integrate, modulate, and interpose a temporal domain in the visual message presented to the RETINAL GANGLION CELLS, with which they synapse in the inner plexiform layer.
Immunologic techniques based on the use of: (1) enzyme-antibody conjugates; (2) enzyme-antigen conjugates; (3) antienzyme antibody followed by its homologous enzyme; or (4) enzyme-antienzyme complexes. These are used histologically for visualizing or labeling tissue specimens.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
The renal tubule portion that extends from the BOWMAN CAPSULE in the KIDNEY CORTEX into the KIDNEY MEDULLA. The proximal tubule consists of a convoluted proximal segment in the cortex, and a distal straight segment descending into the medulla where it forms the U-shaped LOOP OF HENLE.
Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.
The internal portion of the kidney, consisting of striated conical masses, the renal pyramids, whose bases are adjacent to the cortex and whose apices form prominent papillae projecting into the lumen of the minor calyces.
Minute projections of cell membranes which greatly increase the surface area of the cell.
Long convoluted tubules in the nephrons. They collect filtrate from blood passing through the KIDNEY GLOMERULUS and process this filtrate into URINE. Each renal tubule consists of a BOWMAN CAPSULE; PROXIMAL KIDNEY TUBULE; LOOP OF HENLE; DISTAL KIDNEY TUBULE; and KIDNEY COLLECTING DUCT leading to the central cavity of the kidney (KIDNEY PELVIS) that connects to the URETER.
The outer zone of the KIDNEY, beneath the capsule, consisting of KIDNEY GLOMERULUS; KIDNEY TUBULES, DISTAL; and KIDNEY TUBULES, PROXIMAL.
A technique that localizes specific nucleic acid sequences within intact chromosomes, eukaryotic cells, or bacterial cells through the use of specific nucleic acid-labeled probes.
The reconstruction of a continuous two-stranded DNA molecule without mismatch from a molecule which contained damaged regions. The major repair mechanisms are excision repair, in which defective regions in one strand are excised and resynthesized using the complementary base pairing information in the intact strand; photoreactivation repair, in which the lethal and mutagenic effects of ultraviolet light are eliminated; and post-replication repair, in which the primary lesions are not repaired, but the gaps in one daughter duplex are filled in by incorporation of portions of the other (undamaged) daughter duplex. Excision repair and post-replication repair are sometimes referred to as "dark repair" because they do not require light.
Interruptions in the sugar-phosphate backbone of DNA, across both strands adjacently.
Injuries to DNA that introduce deviations from its normal, intact structure and which may, if left unrepaired, result in a MUTATION or a block of DNA REPLICATION. These deviations may be caused by physical or chemical agents and occur by natural or unnatural, introduced circumstances. They include the introduction of illegitimate bases during replication or by deamination or other modification of bases; the loss of a base from the DNA backbone leaving an abasic site; single-strand breaks; double strand breaks; and intrastrand (PYRIMIDINE DIMERS) or interstrand crosslinking. Damage can often be repaired (DNA REPAIR). If the damage is extensive, it can induce APOPTOSIS.
Molecules or ions formed by the incomplete one-electron reduction of oxygen. These reactive oxygen intermediates include SINGLET OXYGEN; SUPEROXIDES; PEROXIDES; HYDROXYL RADICAL; and HYPOCHLOROUS ACID. They contribute to the microbicidal activity of PHAGOCYTES, regulation of signal transduction and gene expression, and the oxidative damage to NUCLEIC ACIDS; PROTEINS; and LIPIDS.
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)
An alkylating agent in cancer therapy that may also act as a mutagen by interfering with and causing damage to DNA.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.

Morphology of intraepithelial corpuscular nerve endings in the nasal respiratory mucosa of the dog. (1/114)

Corpuscular nerve endings in the nasal respiratory mucosa of the dog were investigated by immunohistochemical staining specific for protein gene product 9.5 by light and electron microscopy. In the nasal respiratory mucosa, complex corpuscular endings, which displayed bulbous, laminar and varicose expansions, were distributed on the dorsal elevated part of the nasal septum and on the dorsal nasal concha. The endings were 300-500 microm long and 100-250 microm wide. Some axons gave rise to a single ending while others branched into 2 endings. Cryostat sections revealed that the corpuscular endings were located within the nasal respiratory epithelium. On electron microscopy, immunoreactive nerve terminals that contained organelles, including mitochondria and neurofilaments, were observed within the epithelial layer near the lumen of the nasal cavity. Some terminals contacted the goblet cell. Such terminal regions were covered by the cytoplasmic process of ciliated cells and were never exposed to the lumen of the nasal cavity. These nerve endings are probably activated by pressure changes.  (+info)

Electron diffraction provides new information on human stratum corneum lipid organization studied in relation to depth and temperature. (2/114)

The outermost layer of mammalian skin, the stratum corneum, provides the body with a barrier against transepidermal water loss and penetration of agents from outside. The lipid-rich extracellular matrix surrounding the corneocytes in the stratum corneum is mainly responsible for this barrier function. In this study (cryo-) electron diffraction was applied to obtain information about the local lateral lipid organization in the extracellular matrix in relation to depth in human stratum corneum. For this purpose, stratum corneum grid-strips were prepared from native skin in vivo and ex vivo. It was found that the lipid packing in samples prepared at room temperature is predominantly orthorhombic. In samples prepared at 32 degrees C the presence of a hexagonal packing is more pronounced in the outer layers of the stratum corneum. Gradually increasing the specimen temperature from 30 to 40 degrees C induced a further transition from an orthorhombic to a hexagonal sublattice. At 90 degrees C all lipids were present in a fluid phase. These results are in good agreement with previously reported wide angle X-ray diffraction and Fourier transformed infrared spectroscopy studies. We conclude that the lipids in human stratum corneum are highly ordered throughout the stratum corneum and that electron diffraction allows monitoring of the local lipid organization, which contributes to the understanding of stratum corneum barrier function.  (+info)

Gene delivery to the myocardium by intrapericardial injection. (3/114)

Several studies have demonstrated the feasibility of gene transfer into the heart muscle. However, all the available data also indicate that the extent of transfection remains limited. As an alternative method to intravascular administration, we have developed a novel strategy which uses the pericardial sac. When a replication-deficient adenovirus containing the cDNA encoding a bacterial beta-galactosidase is injected into the pericardial sac of adult Wistar rats the staining is exclusively restricted to the pericardial cell layers. However, injecting a mixture of collagenase and hyaluronidase together with the virus, leads to a large diffusion of the transgene activity, reaching up to 40% of the myocardium. Transgene expression is predominant in the left ventricle and the interventricular septum but limited in the right ventricle. In vivo echocardiographic measurements of the left ventricular diameters at end diastolic and end systolic times show no difference between virus- and sham-injected animals, thus indicating a good clinical tolerance to this strategy of virus delivery. The same protocol has been used with the same efficiency in mice, which leads us to propose injection into the pericardial sac as an effective and harmless method for gene transfer into the heart muscle.  (+info)

Scanning electron microscopic detection of nuclear structures involved in DNA replication. (4/114)

In order to evaluate at the ultrastructural level the three dimensional chromatin arrangement during interphase and particularly during the S phase, the immunogold detection of Bromodeoxyuridine (BrdU), as a marker of DNA synthesis, was performed in human HeLa, HL60, and in murine Friend leukemia cells (FLC). Field emission in lens scanning electron microscopy analysis of ultrathin cryosections revealed the presence of a regular three-dimensional network of fibers in dispersed chromatin. This spatial architecture was apparently constituted mainly of 10 nm filaments organized in loops of about 80-100 nm. Nodal points and the overlapping of such coils appeared as thicker structures of about 30 nm in diameter. Thin filaments of about 5 nm did not show a regular distribution. This three-dimensional fiber organization seemed quite constant in the dispersed chromatin of all the cell lines analyzed. The DNase treatment of the samples selectively removed the 10 nm class fibers, whereas the BrdU labeling confirmed the presence of newly synthesized DNA organized into chromatin units with a regular arrangement. These data suggest that the 10 nm chromatin fiber likely represents the DNA condensation order at which DNA duplication starts and the main weft of a three dimensional network within the interphase nucleus.  (+info)

Pseudorabies virus expressing bovine herpesvirus 1 glycoprotein B exhibits altered neurotropism and increased neurovirulence. (5/114)

Herpesvirus glycoproteins play dominant roles in the initiation of infection of target cells in culture and thus may also influence viral tropism in vivo. Whereas the relative contribution of several nonessential glycoproteins to neurovirulence and neurotropism of Pseudorabies virus (PrV), an alphaherpesvirus which causes Aujeszky's disease in pigs, has recently been uncovered in studies using viral deletion mutants, the importance of essential glycoproteins is more difficult to assess. We isolated an infectious PrV mutant, PrV-9112C2, which lacks the gene encoding the essential PrV glycoprotein B (gB) but stably carries in its genome and expresses the homologous gene of bovine herpesvirus 1 (BHV-1) (A. Kopp and T. C. Mettenleiter, J. Virol. 66:2754-2762, 1992). Apart from exhibiting a slight delay in penetration kinetics, PrV-9112C2 was similar in its growth characteristics in cell culture to wild-type PrV. To analyze the effect of the exchange of these homologous glycoproteins in PrV's natural host, swine, 4-week-old piglets were intranasally infected with 10(6) PFU of either wild-type PrV strain Kaplan (PrV-Ka), PrV-9112C2, or PrV-9112C2R, in which the PrV gB gene was reinserted instead of the BHV-1 gB gene. Animals infected with PrV-Ka and PrV-9112C2R showed a similar course of disease, i.e., high fever, marked respiratory symptoms but minimal neurological disorders, and excretion of high amounts of virus. All animals survived the infection. In contrast, animals infected with PrV-9112C2 showed no respiratory symptoms and developed only mild fever. However, on day 5 after infection, all piglets developed severe central nervous system (CNS) symptoms leading to death within 48 to 72 h. Detailed histological analyses showed that PrV-9112C2R infected all regions of the nasal mucosa and subsequently spread to the CNS preferentially by the trigeminal route. In contrast, PrV-9112C2 primarily infected the olfactory epithelium and spread via the olfactory route. In the CNS, more viral antigen and significantly more pronounced histological changes resulting in more severe encephalitis were found after PrV-9112C2 infection. Thus, our results demonstrate that replacement of PrV gB by the homologous BHV-1 glycoprotein resulted in a dramatic increase in neurovirulence combined with an alteration in the route of neuroinvasion, indicating that the essential gB is involved in determining neurotropism and neurovirulence of PrV.  (+info)

DnaA, the initiator of Escherichia coli chromosomal replication, is located at the cell membrane. (6/114)

Given the lack of a nucleus in prokaryotic cells, the significance of spatial organization in bacterial chromosome replication is only beginning to be fully appreciated. DnaA protein, the initiator of chromosomal replication in Escherichia coli, is purified as a soluble protein, and in vitro it efficiently initiates replication of minichromosomes in membrane-free DNA synthesis reactions. However, its conversion from a replicatively inactive to an active form in vitro occurs through its association with acidic phospholipids in a lipid bilayer. To determine whether the in situ residence of DnaA protein is cytoplasmic, membrane associated, or both, we examined the cellular location of DnaA using immunogold cryothin-section electron microscopy and immunofluorescence. Both of these methods revealed that DnaA is localized at the cell membrane, further suggesting that initiation of chromosomal replication in E. coli is a membrane-affiliated event.  (+info)

Macrocryosectioning of the prostate: a simple technique. (7/114)

Whole mount sections of the prostate are widely used in many laboratories. Macrocryosections of the gland; that is, whole mount frozen sections of the prostate from radical prostatectomies represent a useful new research protocol. The technique is very simple and does not require expensive equipment.  (+info)

Apical accumulation of MARCKS in neural plate cells during neurulation in the chick embryo. (8/114)

BACKGROUND: The neural tube is formed by morphogenetic movements largely dependent on cytoskeletal dynamics. Actin and many of its associated proteins have been proposed as important mediators of neurulation. For instance, mice deficient in MARCKS, an actin cross-linking membrane-associated protein that is regulated by PKC and other kinases, present severe developmental defects, including failure of cranial neural tube closure. RESULTS: To determine the distribution of MARCKS, and its possible relationships with actin during neurulation, chick embryos were transversely sectioned and double labeled with an anti-MARCKS polyclonal antibody and phalloidin. In the neural plate, MARCKS was found ubiquitously distributed at the periphery of the cells, being conspicuously accumulated in the apical cell region, in close proximity to the apical actin meshwork. This asymmetric distribution was particularly noticeable during the bending process. After the closure of the neural tube, the apically accumulated MARCKS disappeared, and this cell region became analogous to the other peripheral cell zones in its MARCKS content. Actin did not display analogous variations, remaining highly concentrated at the cell subapical territory. The transient apical accumulation of MARCKS was found throughout the neural tube axis. The analysis of another epithelial bending movement, during the formation of the lens vesicle, revealed an identical phenomenon. CONCLUSIONS: MARCKS is transiently accumulated at the apical region of neural plate and lens placode cells during processes of bending. This asymmetric subcellular distribution of MARCKS starts before the onset of neural plate bending. These results suggest possible upstream regulatory actions of MARCKS on some functions of the actin subapical meshwork.  (+info)

Leica CM3050 S,The Leica CM3050 S Cryostat Family features superior user comfort with excellent safety standards for practically all types of cryosectioning applications. The Leica CM3050 S is the instrument of choice for all cryosectioning research applications and also for advanced clinical cryosectioning needs,medicine,medical supply,medical supplies,medical product
Visualization of brain monoamine oxidase B (MAO-B) in dementia of Alzheimers type by means of large cryosection autoradiography: a pilot study. - S S Jossan, P G Gillberg, I Karlsson, C G Gottfries, L Oreland
The laboratory and workshop portions of the course provide hands-on introduction to engineering of mouse models, stem cell technologies and tissue analyses. Experimental techniques include genome editing by CRISPR/Cas9, pronuclear microinjection, isolation and culture/manipulation of pre- and post-implantation embryos, embryo transfer, embryo electroporation and roller bottle culture, chimera generation, generation and differentiation of mouse embryonic stem cells and fibroblasts, vibratome and cryosectioning, in situ RNA hybridization, immunostaining, FACS sorting and analysis of hematopoietic stem cells, skeletal preparation, organ explant culture and fluorescent imaging, including live time-lapse microscopy. ...
Im trying to section Xenopus oocytes frozen in OCT with poor results. Lots of cracking and shattering. Im figuring that temp plays a roll here and the little things are very yolky. Is there anybody out in histonetland who may have done some sectioning of frog oocytes or something similar? I actually found a Xenopus histo book but no suggestions for cryosectioning. As is usually the case here, the labs bring the tissue already frozen in OCT. What can I suggest they do prior to or during freezing the make the sections cut better? Does anybody know an optimal temp for cutting frog oocytes? Thanks for any help. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: [email protected]) : ...
We have successfully developed a method for IAA analysis from small numbers of cells of specific cell types using cell sorting of GFP-expressing Arabidopsis lines followed by sensitive MS-based IAA measurements. Using this technique, cell-specific IAA quantification and biosynthesis measurements were possible from lines with GFP expression in as few as four to seven cells per root (pAGL42:GFP, QC cell expressing line) (Nawy et al., 2005). Previously, the only plant tissues in which auxins have been able to be measured at near-cellular resolution are the wood-forming tissues of various tree species, where gradients of IAA concentrations, with maxima in the cambial stem cells, have been observed after cryosectioning of the samples followed by MS analyses (Uggla et al., 1996).. Here, we selected 14 transgenic Arabidopsis lines that collectively express GFP in all cell types of the primary root apex. The lines were selected because they provided the most cell type-specific expression patterns ...
Edwin S. Monuki is the author of these articles in the Journal of Visualized Experiments: Fator-Revestido crescimento Placement Bead em explantes Encéfalo anterior Dorsal, Isolamento do mouse Remoção dorsais Encéfalo anterior, Cultura de rato precursores das células-tronco neurais, Flash Freezing e Cryosectioning E12.5 cérebro de camundongo, Extração de RNA a partir de células Neuroprecursor Usando o Bio-Rad Total de RNA Kit, A genotipagem rápida de tecido Mouse Usando Extract-N-Amp Tissue Sigma PCR Kit
D. Spencer Currle is the author of these articles in the Journal of Visualized Experiments: Flash Freezing e Cryosectioning E12.5 cérebro de camundongo, Cultura de rato precursores das células-tronco neurais, Fator-Revestido crescimento Placement Bead em explantes Encéfalo anterior Dorsal, Rato de isolamento de células Adrenal Cromafim
A single slice of a tomogram of an aldehyde fixed and sucrose infiltrated cryosection with a 3D reconstruction. Erik Bos and Peter J. Peters, Netherlands Cancer Institute, Amsterdam. (see: J. Lefman, P. Zhang, T. Hirai, RM. Weis, J. Juliani, D. Bliss, M. Kessel, E. Bos, P.J. Peters, S. Subramaniam: Three-dimensional electron microscopic imaging of membrane invaginations in Echerichia coli overproducing the chemotaxis receptor Tsr. J. Bacteriol. 2004 Aug; 186(15): 5052-61 ...
The SECOM platform is an integrated solution for correlative light and electron microscopythat enables you to do correlative microscopy extremely fast.
A researcher at LUMC using the THMS600 to look at fluorescently labelled bacteria at liquid nitrogen temperature for correlative light electron microscopy. Professor A.J. Koster and his team at Leiden University Medical Centre are using the THMS600 in a correlative light and electron microscopy setup, to aid the cryo-study of biological specimens.. His group focuses on applications in cell biology, including the study of viral infections and viral replication where fluorescence may be used to pinpoint areas worthy of enhanced investigation. Also of particular interest is the field of vascular biology and the mechanism via which vascular endothelial cells initiate repair in response to injury and inflammation.. His goal is to localize molecular structures in cells using fluorescence microscopy and then transfer the sample to a cryo-electron microscopy (Cryo-EM) set up to image the corresponding macromolecular structures in 3D with nm-scale resolution.. The group wanted a cryo-FM sestup that was ...
TOKYO, Japan, July 21, 2016 - Hitachi High-Technologies Corporation (TSE: 8036, Hitachi High-Tech) and RIKEN, one of Japans national institutes for scientific research, announced today that they have jointly developed MirrorCLEM, a system for simplifying correlative light and electron microscopy (CLEM) which enables the observation of one using both light and scanning electron microscopes (SEM). This new system will be launched for sale by Hitachi High-Tech on 25 July, 2016.. Various types of microscopes are used in a wide variety of fields such as nanotechnology, materials, medical and life sciences. In the medical and life sciences field in particular, SEMs are used to clarify the ultrastructure of cells and tissues, while a type of light microscope called fluorescence microscopes are being used increasingly to observe the localization and behavior of proteins at the molecular level. In recent years, CLEM techniques, which correlate electron microscopy with fluorescence microscopy, have ...
Andrea Falqui. Jul. 30, 2017Science ITO Coating for Correlative Microscopy Correlative microscopy of immunolabeled cells performed by scanning electron and light microscopy needs a suitable coating.
Linkam has been developing cryo stages for correlative microscopy for many years and continues to be at the forefront of cryo correlative microscopy with the latest update of their LINK software for the CMS196M, providing improved imaging capabilities and the new liquid nitrogen autofill system providing longer run times. Click to read more...
Polski) Przypominamy o przetwarzaniu Twoich danych osobowych zgodnych z Rozporządzeniem Parlamentu Europejskiego i Rady (UE) 2016/679 z dnia 27 kwietnia 2016 r. oraz o zasadach, na jakich będzie się to odbywało. Zamykając ten komunikat, zgadzasz się w całości z regulaminem strony ...
High-pressure Freezing. Oct. 26, 2017Science Compression and Crevasses in Vitreous Sections under Different Cutting Conditions Wiley open access: A related research article, again with Jacques
Technology Networks is an internationally recognised publisher that provides access to the latest scientific news, products, research, videos and posters.
The Electron Microscopy facility in the Division of Pathology at the Hospital for Sick Children consists of an EM suite containing three transmission electron microscopes (TEM), a scanning electron microscope (SEM), a field emission SEM (FESEM) with a cryostage and a TEM / scanning transmission electron microscope TEM / STEM. There is a preparative laboratory for routine electron microscopy used primarily for preparing clinical diagnostic specimens as well as preparing specimens for external researchers who wish to do electron microscopy.. In my adjoining laboratory we have all of the preparative equipment required for experimental procedures including, cryofixation for freeze substitution and cryoultramicrotomy and the preparation of thin films. In my laboratory, we have developed novel techniques such as high resolution FESEM secondary electron imaging of tissues and macromolecules in a frozen hydrated state and the ability to image cells, viral particles and large macromolecules in a ...
Sensitive and spatial exploration of the metabolism of tumors at the metabolome level is highly challenging. In this study, we developed an in situ metabolomics method based on ambient mass spectrometry imaging using air flow-assisted desorption electrospray ionization (AFADESI), which can spatially explore the alteration of global metabolites in tissues with high sensitivity. Using this method, we discovered potential histopathological diagnosis biomarkers (including lipids, amino acids, choline, peptides and carnitine) from 52 postoperative lung cancer tissue samples and then subsequently used these biomarkers to generate images for rapid and label-free histopathological diagnosis. These biomarkers were validated with a sensitivity and a specificity of 93.5% and 100%, respectively. Moreover, a single imaging analysis of a cryosection that visualized all these biomarkers, taking tens of minutes, revealed the type and subtype of the cancer. This method could potentially be used as a molecular
For most doctors, any mention of preclinical anatomy teaching evokes memories of the smell of formalin. Medical students in the future, however, are just as likely to use a computer as a scalpel to explore human anatomy, thanks to the Visible Human Project: http://www.nlm.nih.gov/research/visible/visible_human.html. This project, initiated by the US National Library of Medicine, aims to create complete, anatomically detailed, three dimensional representations of male and female human bodies. Transverse computed tomographic, magnetic resonance imaging, and cryosection images have already been collected at 1 mm intervals from a male and a female cadaver-sample images and animations are available on the projects web site ...
The Visible Human Project® is an outgrowth of the NLMs 1986 Long-Range Plan. It is the creation of complete, anatomically detailed, three-dimensional representations of the normal male human body. Cryosection images of representative male cadaver has been completed. The male was sectioned at one millimeter intervals,
Samples (biopsies) taken during surgery to be examined immediately by a cryosection for rapid diagnosis and it can help when resolving about the volume of surgery intervention - in breast surgeries, the genitourinary system, the GI tract, etc. These same samples are subject to a permanent paraffin treatment in regards to providing a precise diagnosis within 2-3 days ...
TY - CHAP. T1 - Correlative light and electron microscopy of autophagosomes. AU - Gudmundsson, Sigurdur Runar. AU - Kahlhofer, Jenny. AU - Baylac, Nastassia. AU - Kallio, Katri Anneli. AU - Eskelinen, Eeva-Liisa. PY - 2019/1. Y1 - 2019/1. N2 - Live-cell imaging has been widely used to study autophagosome biogenesis and maturation. When combined with correlative electron microscopy, this approach can be extended to reveal ultrastructural details in three dimensions. The resolution of electron microscopy is needed when membrane contact sites and tubular connections between organelles are studied.. AB - Live-cell imaging has been widely used to study autophagosome biogenesis and maturation. When combined with correlative electron microscopy, this approach can be extended to reveal ultrastructural details in three dimensions. The resolution of electron microscopy is needed when membrane contact sites and tubular connections between organelles are studied.. KW - 1182 Biochemistry, cell and molecular ...
J.-P. Baudoin, S. Hari, D. Sahmi-Bounsiar, S. Traoré, J. Bou-Khalil, et al.. Correlative light electron microscopy of giant viruses with the SECOM system. New Microbes and New Infections, Wiley Online Library 2018, 26, pp.110-113. ⟨10.1016/j.nmni.2018.09.006⟩. ⟨hal-02006237⟩ ...
Reading Assignment: There are several sections in the Retina and Vitreous section of the BCSC concerning endophthalmitis. Please read through these. In addition, please review the EVS-link attached. ...
Tokuyasu, Naruo/ Shomori, Kohei/ Amano, Kuniki/ Honjo, Soichiro/ Sakamoto, Teruhisa/ Watanabe, Joji/ Amisaki, Masataka/ Morimoto, Masaki/ Uchinaka, Ei/ Yagyu, Takuki/ Saito, Hiroaki/ Ito, Hisao/ Fujiwara, Yoshiyuki/ ...
I agree with Geoff McAuliffe in every detail, you need to perfuse, and you need to fast freeze. If perfusion is new to you, I suggest you review the manual and protocol posted on the left at the following link. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 About the need for fast freezing, see http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freezing%20Artifact.pdf Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 [email protected] http://www.myneurolab.com -----Original Message----- From: [email protected] [mailto:[email protected]] On Behalf Of Anna Elisse Beaudin Sent: Tuesday, December 07, 2004 2:29 PM To: [email protected] Subject: [Histonet] IHC on brain cryosections... help!! Hi all, I am ...
Learn about the newly expanded ZEN technology and new Correlative Microscopy in this interview with John B Yorston, Carl Zeiss. Capabilities of the Sigma VP Field Emission Scanning Electron Microscope are demonstrated and the LSM 700 Scanning Confocal Microscope is shown. These systems combine ultra-resolution with labelling capabilities for a turn-key solution called Shuttle and Find. Filmed by SelectScience at ASCB 2011.
Scanning provides an international and interdisciplinary medium for the rapid exchange of information among all scientists interested in scanning electron, scanning probe, and scanning optical microscopies. Areas of specific interest include all aspects of the instrumentation associated with scanning microscopies, correlative microscopy techniques, stereometry, stereology, analytic techniques, and novel applications of the microscopies.
It is one thing to know that the cryostat will be 30 metres high and 30 metres in diameter; it is another to see these dimensions translated into reality. [...]
To describe characteristics of epiretinal cells at the vitreoretinal interface by correlative light and electron microscopy (CLEM).Epiretinal membrane (ERM) specimens and internal limiting membrane (I...
A 5 nm tomographic slice from a vitreous section of a Saccharomyces cerevisiae cell. (M) is a Mitochondrion and (V) a vacuole. Scale bar, 100 nm. Uppe...
Learn how your ultrastructural investigations can benefit from modern electron microscope techniques. Correlative microscopy provides you deeper insights about cell structure by combination of light and scanning electron microscopy methods.
Visit our booth at FOM 2014 and discover ZEISS systems for light sheet fluorescence microscopy (LSFM), superresolution and correlative microscopy.
Distribution of TRP1 in the Golgi and associated vesicles. (A) Ultrathin cryosections of MNT-1 cells were labeled with anti-TRP1 and PAG 5. TRP1 is often enrich
MR images of the intratemporal portion of the facial nerve were obtained with surface coils using a 0.3-T permanent magnet whole-body imaging system. Various 20FT spin-echo pulse sequences were used to produce 5-mm thick sections with 0.5-mm pixels on a 512 × 512 acquisition matrix. The MR images from normal volunteers were correlated with cryosection specimens of three fresh human cadavers. The seventh nerve was followed in the internal auditory and fallopian canal and through temporal bone to the stylomastoid foramen. The entire labyrinthine, tympanic, and mastoid portions, as well as the geniculate ganglion, could be shown with appropriate scan planes. MR produces excellent images of the facial nerve with high-contrast resolution. Unlike CT, no beam-hardening artifact from the temporal bone is apparent. MR should be a sensitive study for the evaluation of intratemporal facial nerve disease.. ...
The Advanced Light Microscopy Core will be holding a Focus on Microscopy forum on Friday, August 2, 2013. The event will be held at the Jungers Center in Vollum M1441 from 2 to 4:30 p.m. The forum will feature presentations on: New EM Capabilities at OHSU by Chris Arthur, Ph.D., from the FEI Living Lab Devil in the Details: Sample Processing for Electron Superressolution and Correlative Light Microscopy by Danielle Robinson from the Zhong Lab Photoactivated Localization Microscopy with … Read More. ...
Two new and very cool microscopy techniques have been announced recently. One, the optofluidic microscope, could put an entire microscope, including display, into a device the size of an iPod. The other, photoactivated localization microscopy (PALM), was invented by two...
Cryosections of normal and diabetic rat retinas immunostained with anti-LOX primary antibody and rhodamine-conjugated secondary antibody. Blood vessel in the di
Expanding its 3D microcopy portfolio, ZEISS introduced superresolution photoactivated localization microscopy (PALM) in 3D at the Society for Neuroscience Annual Meeting in San Diego, California.
Assessment of different fixation protocols on the presence of membrane-bound vesicles in Caco-2 cells: A multidimensional view by means of correlative light and 3-D transmission electron microscopy, Micron, vol.67, N/A, 2014,pp 20-29 ...
Nature Methods has named Cryo-Electron Microscopy the method of the year 2015. This structural technique, which was always a step behind
Correlative anatomy of the nervous system , Correlative anatomy of the nervous system , کتابخانه الکترونیک و دیجیتال - آذرسا
Individuals with congenital amusia have a lifelong history of unreliable pitch processing. Accordingly, they downweight pitch cues during speech perception and instead rely on other dimensions such as duration. We investigated the neural basis for this strategy. During fMRI, individuals with amusia (N=15) and controls (N=15) read sentences where a comma indicated a grammatical phrase boundary. They then heard two sentences spoken that differed only in pitch and/or duration cues, and selected the best match for the written sentence. Prominent reductions in functional connectivity were detected in the amusia group, between left prefrontal language-related regions and right hemisphere pitch-related regions, which reflected the between-group differences in cue weights in the same groups of listeners. Connectivity differences between these regions were not present during a control task. Our results indicate that the reliability of perceptual dimensions is linked with functional connectivity between ...
TY - JOUR. T1 - The combination of neutrophil-to-lymphocyte ratio and serum carbohydrate antigen 19-9 level as a prognostic indicator in patients with recurrent pancreatic cancer. AU - Sakamoto, Teruhisa. AU - Saito, Hiroaki. AU - Uchinaka, Ei. AU - Morimoto, Masaki. AU - Amisaki, Masataka. AU - Tokuyasu, Naruo. AU - Honjo, Soichiro. AU - Ashida, Keigo. AU - Fujiwara, Yoshiyuki. PY - 2018/9. Y1 - 2018/9. N2 - Background/Aim: We retrospectively investigated the relationship between prognosis and combined neutrophil-to-lymphocyte ratio (NLR) and serum carbohydrate antigen 19-9 (CA19-9) levels in patients with recurrent pancreatic cancer. Patients and Methods: We enrolled 66 patients whose pancreatic cancer recurred. Results: Based on ROC analysis results, the patients were divided into NLRHigh (NLR ≥1.69) or NLRLow (NLR ,1.69), and into CA19-9High (CA19-9 ≥107.95 U/ml) or CA19-9Low (CA19-9 ,107.95 U/ml). When the patients were grouped by combined NLR and CA19-9, their 2-year survival rates ...
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 ...
Kristina Kermanshahche demos Cryo-Electron Microscopy for mapping protein structures on the Intel® Scalable System Framework at Supercomputing 2016.
Modulation of p43 signaling affects early muscle fibrosis. (A,C) Sirius Red staining on cryosections of regenerated Tibialis muscles in wild-type, p43−/− and p43-Tg mice at 28 d.p.i and 70.d.p.i. (B,D) Quantification of Sirius Red staining (H) muscles in wild-type, p43−/− and p43-Tg mice at 28 d.p.i. and 70.d.p.i. (n = 6 for each group). Statistical significance: *p , 0.05; **p , 0.01; ***p , 0.001. Students t-test. Results are expressed as ± sem. ...
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In 1952 Humberto Fernandez Morán introduced cryo ultramicrotomy, which is a similar technique but done at freezing temperatures ... between −20 and −150°C. Cryo ultramicrotomy can be used to cut ultra-thin frozen biological specimens. One of the advantages ...
... cryoultramicrotomy MeSH E05.200.500.620.530.160.260 - frozen sections MeSH E05.200.500.620.620 - replica techniques MeSH ... cryoultramicrotomy MeSH E05.200.750.600.530.160.260 - frozen sections MeSH E05.200.750.600.620 - replica techniques MeSH ...
Leica EM FC7 ‐ Changes your Ultramicrotome Leica EM UC6 or EM UC7 to a Cryoultramicrotome to prepare perfect cryo‐sections for TEM, SEM, AFM & LM.
Cryo-ultramicrotomy is performed after a face is trimmed on the sample. The microtome used has a cryo stage, and cutting of ... Through cryo-ultramicrotomy and heavy metal staining, MVA can image the rubber inclusions with excellent contrast at high ... TEM Analysis of Polymer Rubber Blend Using Cryo-Ultramicrotomy and Osmium Staining. October 6, 2015. by admin ...
Transmission Electron Microscope (TEM) Measurements and Cryo-ultramicrotomy. The bacterial strains used for the TEM ...
Dedicated to the neurobiology of stress in natural populations.. Laboratory at the Arctic Institute of North America, Kluane Lake Base, Yukon and a mobile field laboratory. The following will be available: a complete electrophysiological suite to examine the effects of stress on site, equipment for collection of sensitive brain tissue, and basic services such as an ultra-cold freezer, -20Cº freezer, +4Cº fridge, and an ultrapure water system. ...
Cryofixation and Cryoultramicrotomy. Specimen Collection. Encapsulation. Fixation and Cryoprotection. Coolants. Freezing. ... Subsequent chapters focus on sectioning, cryofixation, and cryoultramicrotomy; positive and negative staining; and the use of ...
Tokuyasu, K. T. Application of cryoultramicrotomy to immunocytochemistry. Journal of microscopy 143, 139-149 (1986). ...
Cryoultramicrotomy. Intestinal Mucosa / cytology, immunology*. Lymphocytes / cytology*. Mice. Tissue Culture Techniques. From ...
The prognostic role of the epidermal growth factor receptor (EGFR) and the related receptor p185HER-2 in lung cancer is as yet undefined. We investigated the immunohistochemical expression of EGFR (monoclonal antibody R1; Amersham) and p185HER-2 (polyclonal antibody A485; Dako) in cryosections. A to …
Cryoultramicrotomy. The technique of using a cryostat or freezing microtome, in which the temperature is regulated to -20 ...
Tokuyasu KT (1989) Use of poly(vinylpyrrolidone) and poly (vinyl alcohol) for cryoultramicrotomy. Histochem J 21:163-171PubMed ... Richter K, Dubochet J (1989) Gluing of vitrified specimens for cryoultramicrotomy. Experientia 45:A42Google Scholar ... Tokuyasu KT (1986) Application of cryoultramicrotomy to immunocytochemistry. J Microsc (Oxford) 143:139-149Google Scholar ... Tsuji S, Anglade P, Daudet-Monsac M, Motelica-Heino I (1992) Cryoultramicrotomy: electrostatic transfer of dry ultrathin frozen ...
Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. TOKUYASU K. T. ...
... cryospecimen preparation by high-pressure freezing and cryoultramicrotomy negative staining and immunogold labeling techniques ...
RMC MT 7000 Ultramicrotome (with cryoultramicrotomy attachment - see below). *RMC MT 7 Ultramicrotome ...
Cryo-ultramicrotomy Lab. available to use at ambient and cryo-temperatures to gather electron transparent sections of a wide ...
Three ultramicrotomes, including one suitable for cryo-ultramicrotomy. *Fume hood, oven for EPON embedding and other TEM sample ...
Dubochet J, McDowall AW (1984a) Cryoultramicrotomy: study of ice crystals and freezing damage. In: Csanady A, Röhlich P, Szabo ...
Immuno-cryoultramicrotomy, p. 71-82. In J. M. Polak and I. M. Varnell (ed.), Immunolabeling for electron microscopy. Elsevier ... The gelatin block with the cells was immersed in cryoprotectant, rapidly frozen before cryoultramicrotomy (UCT with the EM FCS ...
6310118 - Cryoultramicrotomy versus plastic embedding: comparative immunocytochemistry of rat ant.... 18421548 - An f-actin- ...
Tokuyasu, K. T. Application of cryoultramicrotomy to immunocytochemistry. Journal of Microscopy. 143, (Pt 2), 139-149 (1986). ...
In 1952 Humberto Fernandez Morán introduced cryo ultramicrotomy, which is a similar technique but done at freezing temperatures ... between −20 and −150°C. Cryo ultramicrotomy can be used to cut ultra-thin frozen biological specimens. One of the advantages ...
... cryoultramicrotomy MeSH E05.200.500.620.530.160.260 - frozen sections MeSH E05.200.500.620.620 - replica techniques MeSH ... cryoultramicrotomy MeSH E05.200.750.600.530.160.260 - frozen sections MeSH E05.200.750.600.620 - replica techniques MeSH ...
Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem. J. 21:163-171. ...
Tokuyasu KT. Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem J 21: 163-171, 1989. ...
Cryptococcus neoformans cryoultramicrotomy and vesicle fractionation reveals an intimate association between membrane lipids ...
Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem. J. 21:163-171. ...
High-voltage and scanning electron microscopy, and cryo-ultramicrotomy. posted by NITRC Moderator on Dec 16, 2017 ...
Tokuyasu, K. T. (1989). Use of poly(vinylpyrrolidone) and poly(vinylalcohol) for cryoultramicrotomy. Histochem. J. 21, 163-171. ...
Samples were mounted on pins for cryo-ultramicrotomy and frozen in liquid nitrogen. Semithin cryosections (500 nm) for IF ...
1989). Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem. J 21, 163-171. ...
  • A study of proteinases of invasive cells using cryoultramicrotomy and immunogold labelling. (ukzn.ac.za)
  • Includes freeze fracture/freeze etch for SEM, cryoultramicrotomy, freeze substitution, low temperature embedding, and freeze drying of thin samples. (uconn.edu)
  • Electron radiation damage in thin specimens prepared for X-ray microanalysis by cryoultramicrotomy, freeze substitution and normal resin embedding. (faintpower.ga)
  • Cryoultramicrotomy as a preparation method for x-ray microanalysis. (statescale.cf)
  • Although there are still problems to be solved and improvements to be made, we now consider that cryoultramicrotomy has reached the stage of being useful in studying many questions which will not be easily approached otherwise. (nih.gov)
  • Dubochet J, McDowall AW (1984a) Cryoultramicrotomy: study of ice crystals and freezing damage. (springer.com)