scholarly journals A simple and reliable method for correlative light and electron microscopic studies.

1993 ◽  
Vol 41 (5) ◽  
pp. 769-772 ◽  
Author(s):  
J DeFelipe ◽  
A Fairén
Keyword(s):  
Golgi Method ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Simple Method ◽  
Thin Sections ◽  
Axonal Arborization ◽  
Accurate Localization ◽  
Mouse Cerebral Cortex ◽  
Microscopic Studies ◽  
Flat Embedding

We describe in detail a simple method for flat-embedding that can be subsequently used in correlative light and electron microscopic studies. The method can be applied to any material suitable for electron microscopy and is especially useful for study of the synaptology and ultrastructural characteristics of immunocytochemically or morphologically identified neurons or their processes. We present here an example to show how accurately one can delineate the fine details of a complex axonal arborization impregnated with the Golgi method in the mouse cerebral cortex. Golgi-impregnated sections to be studied at the electron microscopic level are osmicated, dehydrated, infiltrated with Araldite resin, flat-embedded, and identified cells or processes photographed. Serial semi-thin sections (1-2 microns thick) are then cut with an ultramicrotome, examined with the light microscope, and the elements rephotographed. Selected semi-thin sections are then resectioned on the ultramicrotome at 60-70 nm and examined electron microscopically. This method allows the systematic and accurate localization of stained cells and processes throughout the successive steps of the procedure.

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.

Correlative immunolocalization with high-resolution light microscopy and Electron Microscopy using London Resin Gold embedded tissue

1995 ◽  
Vol 53 ◽  
pp. 702-703
Author(s):  
M. L. Grove ◽  
B. A. Evans ◽  
D. N. Misra ◽  
J. Zhao ◽  
D. H. Alpers ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Polyclonal Antibodies ◽  
Intracellular Localization ◽  
Intrinsic Factor ◽  
Gastric Epithelium ◽  
Electron Microscopic Level ◽  
Immunoperoxidase Staining ◽  
Rat Tissues ◽  
Electron Microscopic ◽  
Thin Sections

Immunoelectron microscopy specimens are often embedded in hydrophilic resins, like London Resin Gold(LRG), which permit antibody staining without etching (or deplasticizing) the sections. This characteristic of hydrophilic resins also allows for immunohistochemistry at the light level on semi thin sections (0.5μm - 1μm). The ability to do immunohistochemistry at both the light and electron microscopic level on the same tissue block allows focused ultrastructural study. Immunohistochemistry on semi-thin sections displays cellular localization of macromolecules, permitting more specificity in the selection of areas for studying intracellular localization ultrastructurally. We have developed a method for immunoperoxidase staining of LRG embedded tissues, utilizing anti-human polyclonal antibodies directed against intrinsic factor (Fig. 1). Intrinsic factor (IF), a cobalamin binding protein, is known to be produced in the stomach, pancreas and salivary glands of most mammals. We are interested in distribution of IF in gastric epithelium, small intestine (ileum) and supporting tissues in both gastrointestinal tract sites. Previously, we have described the cellular localization of IF in human and rat tissues.

scholarly journals Calmodulin immunoelectron microscopy: redistribution during ram spermatogenesis and epididymal maturation. II.

1986 ◽  
Vol 34 (9) ◽  
pp. 1181-1193 ◽  
Author(s):  
S Weinman ◽  
C Ores-Carton ◽  
F Escaig ◽  
J Feinberg ◽  
S Puszkin
Keyword(s):  
Male Gamete ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Epididymal Maturation ◽  
Coarse Fibers ◽  
Preferential Location ◽  
Lowicryl K4m ◽  
Pleiotropic Regulator ◽  
Monospecific Antibodies

Affinity-purified monospecific antibodies and indirect immunogold and immunoferritin labeling on ultra-thin sections of low-temperature Lowicryl K4M-embedded samples were used to study the redistribution of calmodulin in ram spermatids and epididymal spermatozoa at the electron microscopic level. Calmodulin appeared as an integral component of well-defined structures or organelles of these cells. In young spermatids, calmodulin was localized in the nucleus, cytoplasm, and developing acrosome. During spermatogenesis and epididymal maturation, calmodulin left the acrosome to reach the perinuclear substance and finally became concentrated in the post-acrosomal area of the head, although some calmodulin remained associated with the tip of the acrosome. Such a redistribution is consistent with the preferential location of Ca2+ in the post-acrosomal cytoplasm of ejaculated spermatozoa. Calmodulin was also observed in the flagellum associated with the plasma membrane and with the motility apparatus, between coarse fibers and axonemal microtubules. These changes in calmodulin distribution may account for the Ca2+-dependent regulation of spermatogenesis and sperm maturation. Calmodulin therefore appears to be a pleiotropic regulator of male gamete development and functions.

Comparative ultrastructure of methanogenic bacteria

1975 ◽  
Vol 21 (2) ◽  
pp. 121-129 ◽  
Author(s):  
J. G. Zeikus ◽  
V. G. Bowen
Keyword(s):  
Morphological Characteristics ◽  
Cell Types ◽  
Methanogenic Bacteria ◽  
Diverse Group ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Intracytoplasmic Membranes ◽  
Microscopic Studies ◽  
Different Cell Types ◽  
Comparative Ultrastructure

Electron-microscopic studies using thin sections revealed that methane-producing bacteria were an ultrastructurally diverse group. Fine structure and morphological characteristics separated these bacteria into four discrete cell types. Methanogenic bacteria displayed a gram-positive cell wall that varied considerably among different cell types. Differences in granular inclusions, reserve materials, and intracytoplasmic membranes were observed. Unique ultrastructural features were not shared by all methanogenic species studied.

Electron Microscope Studies on Isolated Plant Protoplasts

1966 ◽  
Vol 21 (6) ◽  
pp. 581-585b ◽  
Author(s):  
E. C. Cocking
Keyword(s):  
Electron Microscope ◽  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Close Association ◽  
Physiological Activity ◽  
Tomato Fruit ◽  
Plant Protoplasts ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Microscopic Studies

Isolated tomato fruit protoplasts have been observed to take up both tobacco mosaic virus and ferritin by the process of pinocytosis. These studies have involved electron microscopic observations on thin sections of suitably fixed and embedded material. These electron microscopic studies have also shown that very close association exists between the nucleus and chloroplasts in these protoplasts and that occasionally there are present channels extending from the plasmalemma into the cytoplasm. The implication of these results is discussed in relation to the general physiological activity of protoplasts.

scholarly journals Conjugated avidin binds to mast cell granules.

1985 ◽  
Vol 33 (1) ◽  
pp. 27-32 ◽  
Author(s):  
M D Tharp ◽  
L L Seelig ◽  
R E Tigelaar ◽  
P R Bergstresser
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Horseradish Peroxidase ◽  
Staining Technique ◽  
Cellular Structures ◽  
Clinical Tool ◽  
Electron Microscopic ◽  
Simple Method ◽  
Human Mast Cells ◽  
Microscopic Studies

The glycoprotein, avidin, conjugated either to the enzyme horseradish peroxidase, or to the fluorochrome dyes, fluorescein or rhodamine, identifies the granules of mast cells in both tissues and cell suspensions. In the absence of prior fixation, mast cells were not identified with conjugated avidin; however, granules released from these cells were stained with this labeled glycoprotein. The specificity of avidin for mast cells was confirmed by the absence of conjugated avidin-positive cells in the skin of mice (S1/S1d) deficient in mature dermal mast cells. Electron microscopic studies confirmed that avidin binds specifically to individual mast cell granules rather than to other cellular structures. Rodent and human mast cells were readily stained with avidin conjugated to horseradish peroxidase or to either of the fluorochrome dyes. The conjugated avidin staining technique is a reliable and simple method for identifying rodent and human mast cells, one that is useful as both an investigative and a clinical tool.

scholarly journals HIGH RESOLUTION AUTORADIOGRAPHY WITH STRIPPING FILM

1971 ◽  
Vol 19 (5) ◽  
pp. 304-309 ◽  
Author(s):  
JOSEPH R. WILLIAMSON ◽  
HANK VAN DEN BOSCH
Keyword(s):  
High Resolution ◽  
Mechanical Device ◽  
Electron Microscopic ◽  
High Quality ◽  
Simple Method ◽  
Thin Sections ◽  
Large Numbers ◽  
High Resolution Autoradiography ◽  
Potential Doubling

A simple method is described for the preparation and use of stripping film of high quality for electron microscopic applications of autoradiography. The advantages of the method are: ( a) uniform monolayers of emulsion are easily prepared with an inexpensive mechanical device and applied to thin sections on conventional grids; ( b) both sides of grids can be coated with emulsion, permitting a potential doubling of sensitivity with, theoretically, no loss of resolution; and ( c) large numbers of grids can be processed simultaneously, assuring identical conditions of development.

Effect of formaldehyde inhalation on rat livers: A light and electron microscopic study

2010 ◽  
Vol 26 (2) ◽  
pp. 113-119 ◽  
Author(s):  
Selman Cikmaz ◽  
Tunc Kutoglu ◽  
Mehmet Kanter ◽  
Recep Mesut
Keyword(s):  
Liver Tissue ◽  
Exposure Period ◽  
Electron Microscopic Level ◽  
Albino Rats ◽  
Electron Microscopic ◽  
Tissue Samples ◽  
Microscopic Evaluation ◽  
Microscopic Studies ◽  
Wistar Albino Rats ◽  
The Individual

It is well known that formaldehyde (FA) is cytotoxic and potentially carcinogenic. Although the individual effects of this reactant on cells has been investigated, the cytotoxicity exerted by the coexistence of FA is poorly understood. The aim of this study was to investigate the effects of FA on the liver in rats, by light and electron microscopic level. We used 18 Wistar albino rats divided into three groups, exposed to 0 (control), 19.7 ppm FA gas for a total of 4 weeks, 8 h/day, 5 days a week (subacute) and 20.3 parts per million (ppm) FA gas for a total of 13 weeks, 8 h/day, 5 days a week (subchronic). After the completion of the exposure period, they were sacrificed by decapitation and their liver tissue samples were taken in order to be processed for light and electron microscopic studies. Light microscopic evaluation of liver tissue samples of FA-exposed rats revealed enlarged sinusoids filled with blood and mononuclear cell infiltration in the portal areas and around the central veins. In addition, some of the hepatocytes showed loss of cytoplasm, and some had a hyperchromatic nucleus. The cells of FA-exposed livers, on the other hand, showed an electron-lucent ground-cytoplasm and a hypertrophy of the smooth-surfaced endoplasmic reticulum. In conclusion, we observed that exposure FA caused diverse histopathological changes indicating the destruction in the liver tissue and this destruction has direct relationship with the length of the exposure period.

Light and Electron Microscopic Studies Regarding Cell Contractility and Cell Coupling in Light Sensitive Smooth Muscle Cells from the Isolated Frog Iris Sphincter

1987 ◽  
Vol 42 (7-8) ◽  
pp. 977-985 ◽  
Author(s):  
Klaus V. Wolf
Keyword(s):  
Gap Junctions ◽  
Freeze Fracture ◽  
Muscle Cells ◽  
Sphincter Muscle ◽  
Cell Coupling ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Cell Contractility ◽  
Microscopic Studies ◽  
Iris Sphincter

(1) In light microscopical studies of living isolated frog irises, it was found that the maximal areas of experimentally light induced contractions in the m. sphincter pupillae were located beneath small illuminated regions. There were no visible contractions of muscle cells outside the illuminated areas. It was shown that exposure to light could directly cause contractions of isolated single sphincter muscle cells. (2) Junctional structures of the iris sphincter cells were studied by means of thin sections and freeze fracture electron microscopy. Intermediate junctions, a few focal tight junctions and occasional small gap junctions were identified. Pit containing intramembranous particles which resemble gap junction connexons were found in large numbers, dispersed over the plasmalemmas of sphincter muscle cells. From these physiological and morphological observations, it is concluded that sphincter muscle cells of the frog iris may be coupled via gap junctions, but that the cell coupling is not sufficiently extensive to form the basis for a functional syncytium.

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