scholarly journals A NEW ELECTRON-DENSE STAIN FOR ELASTIC TISSUE

1970 ◽  
Vol 18 (10) ◽  
pp. 697-708 ◽  
Author(s):  
ERNEST N. ALBERT ◽  
EVERLY FLEISCHER
Keyword(s):  
Electron Microscopy ◽  
Heavy Metals ◽  
Fluorescence Microscopy ◽  
Electron Density ◽  
The Body ◽  
Elastic Tissue ◽  
Electron Microscopic ◽  
Specific Staining ◽  
Microscopic Studies ◽  
Specific Affinity

Tetraphenylporphine sulfonate has been shown to have a specific affinity for elastic tissues of the body in fluorescence microscopy. However, tetraphenylporphine sulfonate does not impart electron density to this tissue and thus is not suitable for electron microscopic studies. Therefore, tetraphenylporphine sulfonate was complexed with various heavy metals in order to use it as a specific stain for elastic tissues in electron microscopy. The silver and gold metallic complexes gave the most consistent and specific staining reactions. These compounds were prepared in this laboratory. The synthesis and staining procedures are described in detail.

A New Electron Dense Stain For Elastin

1969 ◽  
Vol 27 ◽  
pp. 414-415
Author(s):  
Ernest N. Albert
Keyword(s):  
Electron Microscopy ◽  
Heavy Metals ◽  
Phosphotungstic Acid ◽  
Low Ph ◽  
The Body ◽  
Electron Microscopic ◽  
Virtual Absence ◽  
Microscopic Studies ◽  
Specific Affinity ◽  
General Protein

Only phosphotungstic acid (PTA) has been demonstrated to have a reasonably specific affinity for elastin in electron microscopy. However phosphotungstic acid is effective as a specific stain for elastin only at very acid pH. If the pH is increased, PTA is no more a specific stain for elastin, instead it acts as a general protein stain. Therefore at low pH only elastin is visible and one is unable to study the morphological relationships of the elastic components to their surrounding structures due to the virtual absence of contrast in them.Tetraphenylporphine sulfonate (TPPS) has, been shown to have a specific, affinity for elastic tissues of the body in fluorescence microscopy. However TPPS does not impart electron density to this tissue and thus is not suitable for electron microscopic studies. Therefore tetraphenylporphine sulfonate was complexed with various heavy metals in order to use it as a specific stain for elastin in electron microscopy.

Light and scanning electron microscopic studies on Trichodina labrisomi n.sp. from Labrisomas nuchipinnis (Osteichthyes: Labrisomidae) from Mangrove Lake, Bermuda

1993 ◽  
Vol 71 (9) ◽  
pp. 1855-1860
Author(s):  
Thomas G. Rand
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light Microscopy ◽  
Body Shape ◽  
The Body ◽  
Electron Microscopic ◽  
Scanning Electron Microscopic ◽  
Microscopic Studies ◽  
Oral Surface ◽  
Scanning Electron

Trichodina labrisomi n.sp. from the gills of the hairy blenny, Labrisomas nuchipinnis (Quoy and Gaimard), in Mangrove Lake, Bermuda, is described using light microscopy and scanning electron microscopy (SEM). Light microscopy reveals that the dimensions and morphology of T. labrisomi are similar to those given for only 13 previously described species of Trichodina (T. acuta Lom, 1961; T. baicalensis Dogiel, 1957; T. cottomephori Stein, 1979; T. dalli Zhukov, 1964; Trichodina domerguei domerguei Lom and Shtein 1966; T. elegans Stein, 1979; T. fultoni Davis, 1947; T. jadranica Haider, 1964; T. jadranica noblei Lom, 1970; T. partidisci Haider, 1964; T. reticulata Hirschmann et Partsch, 1955; T. tenuidens Faure-Fremiet, 1943; and T. tenuiformis Stein, 1979). However, compared with these similar forms, T. labrisomi n.sp. is distinguished by the dimensions of its body and denticulate ring and (or) the appearance of its silver-impregnated adhesive disc and denticles. Features of T. labrisomi examined by SEM include the body shape, pellicle, aboral pellicular pores, border membrane, aboral ciliary complex, and adoral ciliature, which are described and compared with those of other trichodinids studied using SEM. SEM also revealed that bacilli were distributed circumequatorially over the oral surface of T. labrisomi.

Characterization of Electrophoretic Lipoprotein Fractions: Immunochemical and Electron Microscopic Studies

1972 ◽  
Vol 18 (6) ◽  
pp. 534-538
Author(s):  
Mario Werner ◽  
Albert L Jones
Keyword(s):  
Electron Microscopy ◽  
Particle Size ◽  
Electrophoretic Pattern ◽  
Electron Microscopic ◽  
Lipoprotein Subfractions ◽  
New Techniques ◽  
Microscopic Studies ◽  
Insight Into

Abstract To improve the characterization of electrophoretic lipoprotein subfractions, we developed two new techniques for analyzing lipoproteins after electrophoresis on thin agarose layers. Overlay with antisera exactly localizes specific apoproteins without any distortion caused by antigen diffusion; electron microscopy of eluted fractions determines the varying particle-size distribution. Applied together, these methods can detect individual differences between hyperlipemic samples that are not immediately apparent in the electrophoretic pattern, and should provide valuable new insight into the classification of hyperlipoproteinemias.

scholarly journals Correlating Fluorescence Microscopy with Electron Microscopy

2004 ◽  
Vol 12 (1) ◽  
pp. 3-7
Author(s):  
Stephen W. Carmichael ◽  
Jon Charlesworth
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fluorescence Microscopy ◽  
Fluorescent Probes ◽  
Cell Biology ◽  
Microscopic Image ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Electron Microscopic Image ◽  
Auto Fluorescence

The use of fluorescent probes is becoming more and more common in cell biology. It would be useful if we were able to correlate a fluorescent structure with an electron microscopic image. The ability to definitively identify a fluorescent organelle would be very valuable. Recently, Ying Ren, Michael Kruhlak, and David Bazett-Jones devised a clever technique to correlate a structure visualized in the light microscope, even a fluorescing cell, with transmission electron microscopy (TEM).Two keys to the technique of Ren et al are the use of grids (as used in the TEM) with widely spaced grid bars and the use of Quetol as the embedding resin. The grids allow for cells to be identified between the grid bars, and in turn the bars are used to keep the cell of interest in register throughout the processing for TEM. Quetol resin was used for embedding because of its low auto fluorescence and sectioning properties. The resin also becomes soft and can be cut and easily peeled from glass coverslips when heated to 70°C.

scholarly journals Immuno-electron-microscopic studies on the subcellular distribution of rat liver epoxide hydrolase and the effect of phenobarbitone and 2-acetamidofluorene treatment

1982 ◽  
Vol 202 (3) ◽  
pp. 677-686 ◽  
Author(s):  
F Waechter ◽  
P Bentley ◽  
M Germann ◽  
F Oesch ◽  
W Stäubli
Keyword(s):  
Electron Microscopy ◽  
Rat Liver ◽  
Subcellular Distribution ◽  
Epoxide Hydrolase ◽  
Specific Binding ◽  
Subcellular Fractions ◽  
Electron Microscopic ◽  
Immuno Electron Microscopy ◽  
Microscopic Studies ◽  
The Individual

The distribution of rat liver epoxide hydrolase in various subcellular fractions was investigated by immuno-electron-microscopy. Ferritin-linked monospecific anti-(epoxide hydrolase) immunoglobulins bound specifically to the cytoplasmic surfaces of total microsomal preparations and smooth and rough microsomal fractions as well as the nuclear envelope. Specific binding was not observed when the ferritin conjugates were incubated with peroxisomes, lysosomes and mitochondria. The average specific ferritin load of the individual subcellular fractions correlated well with the measured epoxide hydrolase activities. This correlation was observed with fractions prepared from control, phenobarbitone-treated and 2-acetamidofluorene-treated rats.

scholarly journals Immunogold probes for electron microscopy: evaluation of staining by fluorescence microscopy.

1985 ◽  
Vol 33 (10) ◽  
pp. 995-1000 ◽  
Author(s):  
R B Alexander ◽  
W B Isaacs ◽  
E R Barrack
Keyword(s):  
Electron Microscopy ◽  
Fluorescence Microscopy ◽  
Intermediate Filaments ◽  
Colloidal Gold ◽  
Microscopic Level ◽  
Secondary Antibody ◽  
Light Microscopic Level ◽  
Electron Microscopic ◽  
Cell Monolayers ◽  
Adenocarcinoma Cells

A method is presented whereby the staining of intracellular structures with immunogold probes for electron microscopy can be evaluated at the light microscopic level. Methanol-fixed monolayers of cultured Dunning R-3327-H rat prostatic adenocarcinoma cells were stained for cytokeratins using a two-step immunogold technique consisting of primary anti-keratin antibody followed by gold-labeled secondary antibody. Bound immunogold probe was then visualized with a fluorescent tertiary anti-immunogold probe antibody. Fluorescence microscopy of the whole cell monolayers showed a typical keratin cytoskeleton. The extra staining step did not interfere with subsequent fixation, embedding, and sectioning for electron microscopy, which showed cytoplasmic intermediate filaments decorated with colloidal gold. Using this method, it should be possible to manipulate parameters critical to staining with immunogold probes and to evaluate the labeling without necessitating repeated time-consuming electron microscopic processing. The method also provides a useful correlation between the light microscopic and ultrastructural labeling patterns of immunogold probes.

scholarly journals Using of the method of electron microscopy for the identification and analysis of clinically implicit pathogenetic manifestations

2018 ◽  
Vol 7 (3) ◽  
pp. 113-116
Author(s):  
L. G. Nikonova ◽  
V. V. Banin ◽  
I. G. Stel'nikova
Keyword(s):  
Diabetes Mellitus ◽  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
B Cells ◽  
Secretory Granules ◽  
Electron Microscopic Examination ◽  
The Body ◽  
Electron Microscopic ◽  
Latent Form ◽  
Insulin Cells

Electron microscopic examination of B cells of pancreatic islets of the pancreas in dogs with normal (n=10) and impaired glucose tolerance (n=10) was performed. Ultrastructural features of the organization of insulin cells associated with an increased requirement of the hormone in the body with the latent form of diabetes mellitus are established. In B cells, signs of functional tension due to unregulated secretion, manifested by the expansion of endoplasmic reticulum cisterns, Golgi complex hypertrophy, an increase in the number of immature secretory granules and vacuoles in the cytoplasm are revealed in B cells.

Problems in Myogenesis Terminology in Electron Microscopy

1967 ◽  
Vol 25 ◽  
pp. 38-39
Author(s):  
P.E. Conen ◽  
J.U. Balis ◽  
C.D. Bell
Keyword(s):  
Electron Microscopy ◽  
Cell Types ◽  
Uranyl Acetate ◽  
Electron Microscopic ◽  
Accurate Identification ◽  
Microscopic Studies ◽  
A Cell ◽  
Lead Hydroxide ◽  
Developing Muscle

Myogenesis in man was studied using muscle from 19 fetuses of 8 to 16 weeks gestation which were processed with standard osmium-Epon or glutaraldehyde-osmium-Epon schedules and sections were stained in uranyl acetate and/or lead hydroxide. Particular emphasis was given during this study to presence of basement membrane and myofilaments as additional aids in classification of cell types present in developing muscle.Electron microscopy permits accurate identification of fibroblasts and early cells of muscle series and has been used in studies of myogenesis in chick, and rat. Light microscopy definitions for premyoblasts and myoblasts, and for myocytes at the myotube and muscle fiber stages of development are difficult to apply to electron microscopic studies without modification. For example the term myoblast was used differently by Tello, Katznelson and Boyd to designate a cell destined to become muscle but not recognizable as a muscle cell.

High-Resolution SEM of Colloidal Gold Labels

1988 ◽  
Vol 46 ◽  
pp. 214-217
Author(s):  
Ralph M. Albrecht ◽  
Scott R. Simmons ◽  
James R. Prudent ◽  
Chris M. Erickson
Keyword(s):  
Electron Microscopy ◽  
Colloidal Gold ◽  
Spherical Shape ◽  
Biologically Active ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Color Density ◽  
Microscopic Studies ◽  
Backscattered Electron ◽  
Electron Imaging

Colloidal gold, conjugated to a number of biologically active molecules, including ligand and antibody, provides a useful label for light microscopy and electron microscopy. This stems, in part, from its color, density, and regular spherical shape although the ability to make the particles in a number of defined sizes, the ease of conjugation to biological material, and the retention of activity of bound molecules are also important factors.Although nearly all sizes of colloidal gold particles, from 2.0 nm on up, can be identified in transmission or high voltage transmission electron microscopy, it has generally been the larger sized particles, 15 nm and up, that have proved useful for scanning electron microscopic studies. This is due principally to the resolution limits of conventional SEMs and the need to employ backscattered electron imaging, BEI, to unambiguously define the gold labels.

A Modified Technic of Staining Elastic Tissue for Electron Microscopic Study

1975 ◽  
Vol 33 ◽  
pp. 626-627
Author(s):  
J.A. Nordquist ◽  
K. Chrysant ◽  
A.K. Mandal
Keyword(s):  
Electron Microscopy ◽  
Osmium Tetroxide ◽  
Renal Tissue ◽  
Uranyl Acetate ◽  
Elastic Tissue ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Modified Method ◽  
Tetraphenyl Porphyrin ◽  
Normal Rat

By electron microscopy elastic tissue appear electrolucent in osmium fixed unstained grids as well as grids stained with uranyl acetate and lead citrate (UA + LC). Albert and Fleischer have studied aorta of mice with metalloporphyrins imparting conspicuous electron density to the elastic tissue. We are reporting here a modified method of electron microscopic (EM) study of the elastic tissue using metalloporhyrin, silver tetraphenyl porphyrin sulfonate (STPPS).We have studied the renal arterioles of rats and human in normal and diseased states. Elastic tissue of the aorta from young normal rat served as control for this study. Renal and aortic tissues were fixed in 4 percent glutaraldehyde, post fixed in 1 percent osmium tetroxide and embedded in spurr (blocks). From the blocks of renal tissue, 0.5 μ sections were cut, stained with methylene blue and azure II and studied by light microscopy.

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