Electron microscopic analysis of objects in light microscopic sections

1978 ◽  
Vol 36 (2) ◽  
pp. 80-81
Author(s):  
Arvid B. Maunsbach
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Microscopic Analysis ◽  
Electron Microscopic Level ◽  
Semithin Section ◽  
Cover Glass ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Glass Slides ◽  
Ultrathin Sections

Structural studies in experimental biology or in pathology are frequently extended from the light to the electron microscopic level. This is often done by cutting both semithin (about 1 μm) and thin sections from the same tissue block after embedding for electron microscopy. However, in many studies it would be of great value to analyse the same structure both by light and electron microscopy, i.e. to be able to study by electron microscopy an object which is first detected by light microscopy in a semithin section. To achieve this, a method has been developed by which ultrathin sections are cut directly from the semithin section containing the object of interest.Semithin sections, about 1 μ in thickness, are cut from Epon or Vestopal embedded tissue. The sections are placed on ordinary glass slides and stained with toluidine blue. The sections are studied in the light microscope without a cover glass or mounted in water.

Correlated 3D Light and Electron Microscopy of Large, Complex Structures: Analysis of Transverse Tubules in Heart Failure

1998 ◽  
Vol 4 (S2) ◽  
pp. 440-441
Author(s):  
Maryann E. Martone ◽  
Andrea Thor ◽  
Stephen J. Young ◽  
Mark H. Ellisman.
Keyword(s):  
Electron Microscopy ◽  
Electron Tomography ◽  
Light And Electron Microscopy ◽  
Structural Features ◽  
Electron Microscopic Level ◽  
Specimen Preparation ◽  
Large Sample Size ◽  
Electron Microscopic ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron

Light microscopic imaging has experienced a renaissance in the past decade or so, as new techniques for high resolution 3D light microscopy have become readily available. Light microscopic (LM) analysis of cellular details is desirable in many cases because of the flexibility of staining protocols, the ease of specimen preparation and the relatively large sample size that can be obtained compared to electron microscopic (EM) analysis. Despite these advantages, many light microscopic investigations require additional analysis at the electron microscopic level to resolve fine structural features.High voltage electron microscopy allows the use of relatively thick sections compared to conventional EM and provides the basis for excellent new methods to bridge the gap between microanatomical details revealed by LM and EM methods. When combined with electron tomography, investigators can derive accurate 3D data from these thicker specimens. Through the use of correlated light and electron microscopy, 3D reconstructions of large cellular or subcellular structures can be obtained with the confocal microscope,

Techniques Involved in the Acquisition of Serial Sections of the Atrioventricular Node for Light and Electron Microscopy

1968 ◽  
Vol 26 ◽  
pp. 226-227
Author(s):  
Sheila S. Emmett ◽  
J. C. Thaemert
Keyword(s):  
Electron Microscopy ◽  
Osmium Tetroxide ◽  
Light And Electron Microscopy ◽  
Atrioventricular Node ◽  
Electron Microscopic Level ◽  
Serial Sections ◽  
Electron Microscopic ◽  
Trapezoidal Shape ◽  
Old Mice

The acquisition of serial sections of the atrioventricular node for light and electron microscopy is a formidable task. Ordinary techniques are not adequate if the best possible results are to be achieved at the electron microscopic level. The techniques outlined below have proven to be valuable in locating and determining the position of the AV node.Whole hearts of 2-week old mice were fixed, in situ, by perfusion with 1% phosphate-buffered osmium tetroxide. The hearts were removed from the animals, sectioned transversely into 3 slices approximately equal in thickness, dehydrated in graded concentrations of ethanol and embedded in Epon 812. The block faces were trimmed to a trapezoidal shape ranging in size from 0.75 x 1 mm to 4 x 5 mm. Serial sections approximately 2 microns in thickness were cut with glass knives on a Porter-Blum MT-2 Ultramicrotome. While floating on a drop of water on the knife, each section was stretched with 1 drop of a 1:1, xylene in chloroform mixture applied directly to the section. The sections were picked up individually with a brush, transferred to a glass slide and oven dried for several hours prior to staining.

scholarly journals Light and electron microscopic demonstration of sialic acid residues with the lectin from Limax flavus: a cytochemical affinity technique with the use of fetuin-gold complexes.

1984 ◽  
Vol 32 (11) ◽  
pp. 1167-1176 ◽  
Author(s):  
J Roth ◽  
J M Lucocq ◽  
P M Charest
Keyword(s):  
Electron Microscopy ◽  
Sialic Acid ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
Gold Complexes ◽  
Thin Sections ◽  
Tissue Sections ◽  
Lowicryl K4m ◽  
Hydrolysis Of ◽  
Electron Microscopic Demonstration

The development of a cytochemical affinity technique for the demonstration of sialic acid residues by light and electron microscopy is reported. The lectin from the slug Limax flavus, with its narrow specificity for N-acetyl- and N-glycolylneuraminic acid, was applied to tissue sections. Subsequently fetuin-gold complexes were used to visualize the tissue-bound lectin. Different cytochemical controls, including sugar inhibition tests, neuraminidase digestion, the use of fetuin-gold complexes alone, or acid hydrolysis of sections, proved the specificity of the technique. Postembedding staining was performed on frozen, paraffin, or semithin resin sections for light microscopy and on thin sections from low temperature Lowicryl K4M-embedded material for electron microscopy. The distribution of sialic acid residues in rat pancreas, liver, and colonic mucosa was investigated.

scholarly journals Fine Structural Specific Visualization of RNA on Ultrathin Sections

1998 ◽  
Vol 46 (3) ◽  
pp. 389-395 ◽  
Author(s):  
Marco Biggiogera ◽  
Stanislav Fakan
Keyword(s):  
Rnase A ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Acrylic Resins ◽  
Cell Components ◽  
Ultrathin Sections ◽  
A New Technique ◽  
Interchromatin Granules ◽  
Perichromatin Granules

We describe a new technique that allows specific visualization of RNA at the electron microscopic level by means of terbium citrate. Under the conditions presented here, terbium binds selectively to RNA and stains nucleoli, interchromatin granules, perichromatin fibrils, perichromatin granules, and coiled bodies in the cell nucleus, whereas ribosomes are the only contrasted structures in the cytoplasm. All the cell components contrasted by terbium are known to contain RNA. When ultrathin sections are pretreated with RNase A or nuclease S1 (specific for single-stranded nucleic acids), staining does not occur. Neither DNase nor pronase influences the reaction. We conclude that terbium staining is selective for RNA and especially for single-stranded RNA. The staining can be performed on thin sections of material embedded both in epoxy and in acrylic resins. The technique is not influenced by the aldehyde fixative used and can also be utilized after immunolabeling. The endproduct is very fine and, although weak in contrast, is suitable for high-resolution observations.

mRNA localization by Electron microscopic in situ hybridization

1991 ◽  
Vol 49 ◽  
pp. 430-431
Author(s):  
J. A. Pollock ◽  
M. Martone ◽  
T. Deerinck ◽  
M. H. Ellisman
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Nucleic Acids ◽  
Recombinant Dna ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
High Voltage Electron Microscopy ◽  
Functional Sites ◽  
Voltage Electron

Localization of specific proteins in cells by both light and electron microscopy has been facilitate by the availability of antibodies that recognize unique features of these proteins. High resolution localization studies conducted over the last 25 years have allowed biologists to study the synthesis, translocation and ultimate functional sites for many important classes of proteins. Recently, recombinant DNA techniques in molecular biology have allowed the production of specific probes for localization of nucleic acids by “in situ” hybridization. The availability of these probes potentially opens a new set of questions to experimental investigation regarding the subcellular distribution of specific DNA's and RNA's. Nucleic acids have a much lower “copy number” per cell than a typical protein, ranging from one copy to perhaps several thousand. Therefore, sensitive, high resolution techniques are required. There are several reasons why Intermediate Voltage Electron Microscopy (IVEM) and High Voltage Electron Microscopy (HVEM) are most useful for localization of nucleic acids in situ.

Electron Microscopic Analysis of Myonecrosis Induced by a Pure Component Isolated From Rattlesnake Venom

1976 ◽  
Vol 34 ◽  
pp. 292-293
Author(s):  
Charlotte L. Ownby ◽  
David Cameron ◽  
Anthony T. Tu
Keyword(s):  
Gel Filtration ◽  
Microscopic Analysis ◽  
The United States ◽  
Exchange Chromatography ◽  
Pure Component ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Mouse Muscle ◽  
Major Health Problem ◽  
Rattlesnake Venom

In the United States the major health problem resulting from snakebite poisoning is local tissue damage, i.e. hemorrhage and myonecrosis. Since commercial antivenin does not usually prevent such damage to tissue, a more effective treatment of snakebite-induced myonecrosis is needed. To aid in the development of such a treatment the pathogenesis of myonecrosis induced by a pure component of rattlesnake venom was studied at the electron microscopic level.The pure component, a small (4,300 mol. wt.), basic (isoelectric point of 9.6) protein, was isolated from crude prairie rattlesnake (Crotalus viridis viridis) venom by gel filtration (Sephadex G-50) followed by cation exchange chromatography (Sephadex C-25), and shown to be pure by electrophoresis. Selection of the myotoxic component was based on light microscopic observations of injected mouse muscle.

Electron Microscopic Characterization and Quantitation of Air Borne Particulate Matter with Special Emphasis on Asbestos

1972 ◽  
Vol 30 ◽  
pp. 356-357
Author(s):  
Peter K. Mueller ◽  
Glenn R. Smith ◽  
Leslie M Carpenter ◽  
Ronald L. Stanley
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Ambient Air ◽  
Quality Standard ◽  
Primary Objective ◽  
Cellulose Ester ◽  
Main Concept ◽  
Electron Microscopic ◽  
Air Samples ◽  
Diffraction Patterns

At the present time the primary objective of the electron microscopy group of the Air and Industrial Hygiene Laboratory is the development of a method suitable for use in establishing an air quality standard for asbestos in ambient air and for use in its surveillance. The main concept and thrust of our approach for the development of this method is to obtain a true picture of fiber occurrence as a function of particle size and asbestos type utilizing light and electron microscopy.We have now available an electron micrographic atlas of all asbestos types including selected area diffraction patterns and examples of fibers isolated from air samples. Several alternative approaches for measuring asbestos in ambient air have been developed and/or evaluated. Our experiences in this regard will be described. The most promising method involves: 1) taking air samples on cellulose ester membrane filters with a nominal pore size of 0.8 micron; 2) ashing in a low temperature oxygen plasma for several hours;

scholarly journals FIXATION OF NEURAL TISSUES FOR ELECTRON MICROSCOPY BY PERFUSION WITH SOLUTIONS OF OSMIUM TETROXIDE

1962 ◽  
Vol 12 (2) ◽  
pp. 385-410 ◽  
Author(s):  
Sanford L. Palay ◽  
S. M. McGee-Russell ◽  
Spencer Gordon ◽  
Mary A. Grillo
Keyword(s):  
Electron Microscopy ◽  
Nervous System ◽  
Osmium Tetroxide ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Vascular Perfusion ◽  
Neural Tissues ◽  
Structural Relations ◽  
Cellular Components

This paper describes in detail a method for obtaining nearly uniform fixation of the nervous system by vascular perfusion with solutions of osmium tetroxide. Criteria are given for evaluating the degree of success achieved in the preservation of all the cellular components of the nervous system. The method permits analysis of the structural relations between cells at the electron microscopic level to an extent that has not been possible heretofore.

Phase-contrast and electron-microscopic observations on a membranous complex in primary spermatocytes of the mouse

1975 ◽  
Vol 18 (1) ◽  
pp. 1-17
Author(s):  
A. Pleshkewych ◽  
L. Levine
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Phase Contrast ◽  
Cultured Cells ◽  
Light And Electron Microscopy ◽  
Cytoplasmic Inclusion ◽  
Electron Microscopic ◽  
Secondary Spermatocyte ◽  
Living Mouse ◽  
Circular Contour

A prominent cytoplasmic inclusion present in living mouse primary spermatocytes has been observed by both light and electron microscopy. It began to form at prometaphase and continued to increase in thickness and length as the cells developed. By metaphase it was a distinct sausage-shaped boundary that enclosed a portion of the cytoplasm between the spindle and the cell membrane. At the end of metaphase, the inclusion reached its maximum length. At telophase, it was divided between the daughter secondaries. The inclusion persisted as a circular contour in the interphase secondary spermatocyte. Electron microscopy of the same cultured cells that were previously observed with light microscopy revealed that the inclusion was a distinctive formation of membranes. It consisted of agranular cisternae and vesicles, and was therefore a membranous complex. Many of the smaller vesicles in the membranous complex resembled those found in the spindle. The cisternae in the membranous complex were identical to the cisternal endoplasmic reticulum of interphase primary spermatocytes. Nevertheless, the organization of vesicles and cisternae into the membranous complex was unique for the primaries in division stages, since such an organization was not present in their interphase stages.

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