Staining methods for sections of epon-embedded tissues for light microscopy

1970 ◽  
Vol 48 (1) ◽  
pp. 189-191 ◽  
Author(s):  
C. Roland Leeson ◽  
Thomas S. Leeson
Keyword(s):  
Light Microscopy ◽  
Synthetic Resin ◽  
Electron Microscopic ◽  
Glass Slides ◽  
Staining Methods

Sections 0.5–2 μ thick are mounted on clean glass slides and allowed to dry. A number of staining procedures are described. After the sections are stained, permanent preparations are made by mounting them in a synthetic resin. The methods result in sections which are suitable for routine light microscopy and for comparison with adjacent electron microscopic sections.

Cytochemical methods for the rapid demonstration of infection at the tissue-biomaterial interface

1992 ◽  
Vol 50 (1) ◽  
pp. 634-635
Author(s):  
J. Hanker ◽  
J.J. Dobbins ◽  
P.E. Yates ◽  
B.L. Giammara
Keyword(s):  
Light Microscopy ◽  
Culture Techniques ◽  
The Past ◽  
Glass Slides ◽  
Staining Methods ◽  
The Times

Infection can be an extensive problem developing during implantation of an autogenous or prosthetic (biomaterial) device or graft. Although culture techniques are invaluable in identifying the responsible microorganisms, the times required frequently emphasize the need for rapid staining methods which can reduce the classification times from days to hours. Studies in our laboratories over the past few years have resulted in microwave-accelerated stains and in methods developed by Giammara, which enable the rapid study of glass slides and coverslips by electron as well as light microscopy.

Immunocytochemistry. A Review of Methodology for Electron Microscopic Applicaiton

1974 ◽  
Vol 32 ◽  
pp. 328-329
Author(s):  
Joseph E. Mazurkiewicz
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Biological Interest ◽  
Investigative Approach ◽  
Immunologic Activity ◽  
Dense Label

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.

A Method for Electron Microscopic Study of Tissue Culture Cell Monolayers Grown in Tubes

1967 ◽  
Vol 25 ◽  
pp. 132-133
Author(s):  
R. Stephens ◽  
G. Schidlovsky ◽  
S. Kuzmic ◽  
P. Gaudreau
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Light Microscopy ◽  
Cell Sheet ◽  
Culture Cell ◽  
Tissue Culture Cell ◽  
Electron Microscopic ◽  
Cell Sheets ◽  
Morphological Alterations ◽  
Culture Cells

The usual method of scraping or trypsinization to detach tissue culture cell sheets from their glass substrate for further pelletization and processing for electron microscopy introduces objectionable morphological alterations. It is also impossible under these conditions to study a particular area or individual cell which have been preselected by light microscopy in the living state.Several schemes which obviate centrifugation and allow the embedding of nondetached tissue culture cells have been proposed. However, they all preserve only a small part of the cell sheet and make use of inverted gelatin capsules which are in this case difficult to handle.We have evolved and used over a period of several years a technique which allows the embedding of a complete cell sheet growing at the inner surface of a tissue culture roller tube. Observation of the same cell by light microscopy in the living and embedded states followed by electron microscopy is performed conveniently.

Correlative video enhanced differential interference contrast light microscopy (VDIC)-LM), low-voltage high-resolution SEM (LV-HR-SEM), and high-voltage electron microscopy (HVEM) in the observation of gold-labelled surface antigens and the subjacent cytoskeletal matrix

1989 ◽  
Vol 47 ◽  
pp. 904-905
Author(s):  
Ralph M. Albrecht ◽  
Scott R. Simmons ◽  
Marek Malecki
Keyword(s):  
High Resolution ◽  
Light Microscopy ◽  
Low Voltage ◽  
Microscopic Analysis ◽  
Cell Labelling ◽  
Video Technology ◽  
Electron Microscopic ◽  
Image Capture ◽  
High Voltage Electron Microscopy ◽  
Fluorescence Studies

The development of video-enhanced light microscopy (LM) as well as associated image processing and analysis have significantly broadened the scope of investigations which can be undertaken using (LM). Interference/polarization based microscopies can provide high resolution and higher levels of “detectability” especially in unstained living systems. Confocal light microscopy also holds the promise of further improvements in resolution, fluorescence studies, and 3 dimensional reconstruction. Video technology now provides, among other things, a means to detect differences in contrast difficult to detect with the human eye; furthermore, computerized image capture, processing, and analysis can be used to enhance features of interest, average images, subtract background, and provide a quantitative basis to studies of cells, cell features, cell labelling, and so forth. Improvements in video technology, image capture, and cost-effective computer image analysis/processing have contributed to the utility and potential of the various interference and confocal microscopic instrumentation.Electron microscopic technology has made advances as well. Microprocessor control and improved design have contributed to high resolution SEMs which have imaging capability at the molecular level and can operate at a range of accelerating voltages starting at 1KV. Improvements have also been seen in the HVEM and IVEM transmission instruments. As a whole, these advances in LM and EM microscopic technology provide the biologist with an array of information on structure, composition, and function which can be obtained from a single specimen. Corrrelative light microscopic analysis permits examination of living specimens and is critical where the “history” of a cell, cellular components, or labels needs to be known up to the time of chemical or physical fixation. Features such as cytoskeletal elements or gold label as small as 0.01 μm, well below the 0.2 μm limits of LM resolution, can be “detected” and their movement followed by VDIC-LM. Appropriate identification and preparation can then lead to the examination of surface detail and surface label with stereo LV-HR-SEM. Increasing the KV in the HR-SEM while viewing uncoated or thinly coated specimens can provide information from beneath the surface as well as increasing Z contrast so that positive identification of surface and subsurface colloidal gold or other heavy metal labelled/stained material is possible. Further examination of the same cells using stereo HVEM or IVEM provides information on internal ultrastructure and on the relationship of labelled material to cytoskeletal or organellar distribution, A wide variety of investigations can benefit from this correlative approach and a number of instrumentational configurations and preparative pathways can be tailored for the particular study. For a surprisingly small investment in time and technique, it is often possible to clear ambiguities or questions that arise when a finding is presented in the context of only one modality.

scholarly journals THE GLOMERULUS IN EXPERIMENTAL RENAL DISEASE IN RATS AS OBSERVED BY LIGHT AND ELECTRON MICROSCOPY

1955 ◽  
Vol 102 (5) ◽  
pp. 573-580 ◽  
Author(s):  
Carolyn F. Piel ◽  
Luther Dong ◽  
F.W.S. Modern ◽  
Joseph R. Goodman ◽  
Roger Moore
Keyword(s):  
Electron Microscopy ◽  
Endothelial Cells ◽  
Basement Membrane ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
Crescent Formation ◽  
Colloidal Iron ◽  
Narrow Channels ◽  
Glomerular Capillary

Nephrotoxic serum disease in rats has been studied by light and electron microscopy from 1 hour to 10 weeks after production of the disease. By light microscopy leucocytic infiltration of the glomerular capillary was observed between the 3rd and 6th hour. At 6 hours an increase in colloidal iron-positive material was observed coating the extraluminal surface of the capillaries. Also at this time swelling of the endothelial cells becomes prominent. By 72 hours, thickening of the basement membrane was observed. Glomerular capillary thrombi were observed in approximately half the tissue examined in the first 2 weeks of disease. 50 per cent of the animals showed severe chronic lesions, exudation into the capsular space, crescent formation, and obliteration of glomeruli. At 1 hour electron microscopic pictures showed that osmophilic material may line the foot processes of the epithelial cells and obliterate all but narrow channels of the space between the feet. By 6 hours thickening of the basement membrane was prominent. This change persisted throughout 10 weeks of observation. The tissue from animals which had severe chronic alterations by light microscopy revealed changes which could not be interpreted at this time.

The Evaluation of Ciliary Function: Electron versus Light Microscopy

1998 ◽  
Vol 12 (3) ◽  
pp. 199-202 ◽  
Author(s):  
Stephen B. Kupferberg ◽  
John P. Bent ◽  
Edward S. Porubsky
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Primary Ciliary Dyskinesia ◽  
Imaging Techniques ◽  
Electron Microscopic Examination ◽  
Definitive Diagnosis ◽  
Electron Microscopic ◽  
Ciliary Function ◽  
Physical Findings ◽  
Ciliary Dyskinesia

Diagnosing Primary Ciliary Dyskinesia can often be difficult. Physical findings suggest the disease, but definitive diagnosis should be made with a ciliary biopsy. Twenty biopsies were obtained from 16 patients and all underwent both light and electron microscopic examination. In 8/20 (40%) there was a discrepancy between the different imaging techniques. Therefore, light microscopy should be used to assess adequacy of biopsy and motion of the cilia along with electron microscopy to examine ultrastructure.

FINE STRUCTURE OF NUCLEOLI AND RNA-CONTAINING CHROMOSOME REGIONS OF SALIVARY GLAND CHROMOSOMES OF CHIRONOMIDS AND THEIR INTERRELATIONSHIP

1964 ◽  
Vol 42 (6) ◽  
pp. 1147-1155 ◽  
Author(s):  
V. I. Kalnins ◽  
H. F. Stich ◽  
S. A. Bencosme
Keyword(s):  
Fine Structure ◽  
Salivary Gland ◽  
Electron Microscope ◽  
Light Microscopy ◽  
Messenger Rna ◽  
Balbiani Ring ◽  
Electron Microscopic ◽  
Chironomid Species ◽  
Balbiani Rings ◽  
Chromosome Regions

Electron microscope studies of salivary gland nuclei of four chironomid species have shown that the RNA-containing chromosome regions and associated structures, which by light microscopy exhibit a great variety of structures such as bands, granules, micronucleoli, nucleoli, puffs, and Balbiani rings, consist of only few basic units: pars amorpha, nucleolonema, and Balbiani ring granules. The fine structure of the nucleoli and spherical micronucleoli located at various chromosome regions appears to be identical, consisting of pars amorpha, which contains fibers of varying diameters, and strands of nucleolonema composed of fibers and ribosome-like granules. The arrangement of pars amorpha and nucleolonema of nucleoli and spherical micronucleoli follows a consistent pattern. Chromosome fibers are closely associated with pars amorpha, whereas strands of nucleolonema border only the surfaces of pars amorpha. Balbiani ring granules, which have a diameter of 300 Å to 500 Å and are characterized by a particular structure, accumulate in Balbiani rings, in many chromosome regions, and in nuclear sap. In the Balbiani ring these granules seem to be attached to 100 Å chromosome fibers. They are absent in nucleoli and micronucleoli. The possible correlation between our electron microscopic observations and the present-day concept of ribosomal and messenger RNA production is discussed.

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