scholarly journals FURTHER ELECTRON MICROSCOPE STUDIES ON FIBRILLAR ORGANIZATION OF THE GROUND CYTOPLASM OF CHAOS CHAOS

1968 ◽  
Vol 38 (1) ◽  
pp. 40-50 ◽  
Author(s):  
Vivianne T. Nachmias
Keyword(s):  
Electron Microscope ◽  
Alcian Blue ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Negative Staining ◽  
Glutaraldehyde Fixation

Further evidence for fibrillar organization of the ground cytoplasm of Chaos chaos is presented. Fixations with osmium tetroxide at pH 6 or 8 and with glutaraldehyde at pH 6 or 7 were used on two preparations: (a) single actively streaming cells; (b) prechilled cells treated with 0.05% Alcian blue in the cold and returned to room temperature for 5–10 min. In addition, a 50,000 g pellet of homogenized cells was examined after fixation with glutaraldehyde-formaldehyde alone. In sections from actively streaming cells considerable numbers of filaments were observed in the uroid regions after glutaraldehyde fixation, whereas only traces of filaments were seen after osmium tetroxide fixation at either pH 6 or 8. Microtubules were not seen. In sections from dye-treated cells, filaments (4–6 mµ) and fibrils (12–15 mµ) were found with all three fixatives. The 50,000 g pellet was heterogeneous but contained both clumps of fibrils and single thick fibrils like those seen in the cytoplasm of dye-treated cells. Many fibrils of the same dimensions (12–15 mµ wide, 0.5 µ long) were also seen in the supernatant above the pellet. Negative staining showed that some fibrils separated into at least three strands of 4–6 mµ filaments.

Observations of thick and thin sections in a field-emission SEM

1994 ◽  
Vol 52 ◽  
pp. 1018-1019
Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley
Keyword(s):  
Electron Microscope ◽  
Field Emission ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Transmission Electron ◽  
Conventional Transmission Electron Microscope ◽  
Scanning Electron

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.

Lipid in Erysiphe graminis hordei and its possible role during germination

1970 ◽  
Vol 16 (11) ◽  
pp. 1041-1044 ◽  
Author(s):  
W. E. McKeen
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Erysiphe Graminis ◽  
Glutaraldehyde Fixation ◽  
Sudan Black B ◽  
Erysiphe Graminis Hordei ◽  
Sudan Iv ◽  
Mother Cells ◽  
Hyphal Tip

Osmiophilic bodies appear in parts of the colonial growth of Erysiphe graminis DC. f. sp. hordei Em Marchal culture CR3 growing on the susceptible commercial Keystone variety of barley. They are readily observed by the light and electron microscope after osmium tetroxide staining and are abundant in conidiophores, conidia, and mycelium except in haustorial mother cells, in which they are usually absent. The metabolism of haustorial mother cells is distinct and the fine structure of adjoining cells is frequently different. Osmiophilic bodies are absent from the growing hyphal tip, but gradually increase in number and size further back in the terminal cell. Electron micrographs show that they are intracytoplasmic, intravacuolar, and up to 1 μ in diameter. When the colony is washed with acetone or alcohol rather than with aqueous buffer, after glutaraldehyde fixation, before osmium tetroxide fixation, the osmiophilic bodies are removed, indicating that they are lipids. Fat stains, Sudan black B, and Sudan IV stain these bodies. Perhaps the water needs of the germinating conidium are met in part by the oxidation of fats.

scholarly journals PRESERVATION OF MYELIN LAMELLAR STRUCTURE IN THE ABSENCE OF LIPID

1967 ◽  
Vol 34 (3) ◽  
pp. 817-826 ◽  
Author(s):  
Leonard Napolitano ◽  
Francis Lebaron ◽  
Joseph Scaletti
Keyword(s):  
Fatty Acids ◽  
Liquid Chromatography ◽  
Fine Structure ◽  
Electron Microscope ◽  
Lamellar Structure ◽  
Osmium Tetroxide ◽  
Gas Liquid Chromatography ◽  
Glutaraldehyde Fixation ◽  
Thin Sections ◽  
Sciatic Nerves

The fine structure of myelin was studied in glutaraldehyde-fixed rat sciatic nerves depleted of lipid by acetone, chloroform:methanol (2:1 v/v), and chloroform:methanol:concentrated HCl (200:100:1, v/v/v). One portion of each of these nerves, plus the extracts, was saponified and analyzed by gas-liquid chromatography for fatty acids. The remainder of each nerve was stained in osmium tetroxide in CCl4 (5g/100cc) and was embedded in Epon 812. Thin sections, examined in the electron microscope, revealed the preservation of myelin lamellar structure with a 170 A periodicity in nerves depleted of 98% of their lipids. Preservation of myelin lamellar structure depended on glutaraldehyde fixation and the introduction of osmium tetroxide in a nonpolar vehicle (CCl4) after the lipids had been extracted. It is concluded that the periodic lamellar structure in electron micrographs of myelin depleted of lipid results from the complexing of osmium tetroxide, plus uranyl and lead stains, with protein.

Mesosome-like Structures in Cryptococcus neoformans Studied by Chemical Fixation and by the Freeze-etching Technique

1968 ◽  
Vol 26 ◽  
pp. 80-81
Author(s):  
M. R. Edwards ◽  
S. C. Holt
Keyword(s):  
Electron Microscope ◽  
Cryptococcus Neoformans ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Chemical Fixation ◽  
Etching Technique ◽  
Pathogenic Yeast ◽  
Thin Sections ◽  
Lead Salts ◽  
Freeze Etching

The general features of Cryptococcus neoformans, a pathogenic yeast, have been studied with the electron microscope. In the course of such a study it was noted that the plasma membrane of C. neoformans, occasionally invaginated into the cytoplasm and formed membranous organelles which resembled bacterial mesosomes. The present investigation was undertaken in order to examine such structures in detail and to compare the results from chemical fixation with those of freeze-etching.Cells were grown in Sabouraud's agar at 25-27° C for 24-48 hr and fixed with 4% glutaraldehyde in 0.15 M phosphate (Sbrensen's) buffer, at room temperature, for 2 hr; after being thoroughly washed in the buffer and post-fixed in osmium tetroxide, in the same buffer, they were dehydrated in ethyl alcohol and embedded in Epon. Thin sections were cut in a LKB microtome, double stained with uranyl and lead salts and examined in the Siemens Elmiskop IA.

scholarly journals Ultrastructure of microfilament bundles in baby hamster kidney (BHK-21) cells. The use of tannic acid.

1979 ◽  
Vol 80 (3) ◽  
pp. 759-766 ◽  
Author(s):  
R D Goldman ◽  
B Chojnacki ◽  
M J Yerna
Keyword(s):  
Electron Microscope ◽  
Electron Density ◽  
Tannic Acid ◽  
Osmium Tetroxide ◽  
Stress Fibers ◽  
Supramolecular Organization ◽  
Glutaraldehyde Fixation ◽  
Microfilament Bundles ◽  
Electron Density Shifts

After standard glutaraldehyde-osmium tetroxide fixation procedures, the majority of microfilament bundles in BHK-21 cells exhibit relatively uniform electron density along their long axes. The inclusion of tannic acid in the glutaraldehyde fixation solution results in obvious electron density shifts along the majority of microfilament bundles. Striated patterens are frequently observed which consist of regularly spaced electron dense (D) and electron lucid (L) bands. A striated pattern is also observed along many BHK-21 stress fibers after processing for indirect immunofluorescence utilizing BHK-21 myosin antiserum. A direct correlation of these periodicities seen by light and electron microscope techniques is impossible at the present time. However, comparative measurements indicate that the overall patterns seen in the immunofluorescence and electron microscope preparations are similar. The ultrastructural results provide an initial clue for the ultimate determination of the supramolecular organization of contracile proteins other than actin within the microfilament bundles of non-muscle cells.

A Liquid Nitrogen Cooled Stage for STEM

1974 ◽  
Vol 32 ◽  
pp. 444-445
Author(s):  
Louis T. Germinario
Keyword(s):  
Electron Microscope ◽  
High Resolution ◽  
Liquid Nitrogen ◽  
Room Temperature ◽  
Objective Lens ◽  
Thermal Drift ◽  
Specimen Holder ◽  
Resolution Capability ◽  
Focal Position

A liquid nitrogen stage has been developed for the JEOL JEM-100B electron microscope equipped with a scanning attachment. The design is a modification of the standard JEM-100B SEM specimen holder with specimen cooling to any temperatures In the range ~ 55°K to room temperature. Since the specimen plane is maintained at the ‘high resolution’ focal position of the objective lens and ‘bumping’ and thermal drift la minimized by supercooling the liquid nitrogen, the high resolution capability of the microscope is maintained (Fig.4).

Further Observations on the Virus of Epizootic Diarrhea of Infant Mice. An Electron Microscopic Study

1967 ◽  
Vol 25 ◽  
pp. 110-111
Author(s):  
W. G. Banfield ◽  
G. Kasnic ◽  
J. H. Blackwell
Keyword(s):  
Electron Microscope ◽  
Infected Cell ◽  
Microscopic Study ◽  
Intestinal Epithelium ◽  
Ultrastructural Study ◽  
Osmium Tetroxide ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Virus Particles ◽  
Infected Cells

An ultrastructural study of the intestinal epithelium of mice infected with the agent of epizootic diarrhea of infant mice (EDIM virus) was first performed by Adams and Kraft. We have extended their observations and have found developmental forms of the virus and associated structures not reported by them.Three-day-old NLM strain mice were infected with EDIM virus and killed 48 to 168 hours later. Specimens of bowel were fixed in glutaraldehyde, post fixed in osmium tetroxide and embedded in epon. Sections were stained with uranyl magnesium acetate followed by lead citrate and examined in an updated RCA EMU-3F electron microscope.The cells containing virus particles (infected) are at the tips of the villi and occur throughout the intestine from duodenum through colon. All developmental forms of the virus are present from 48 to 168 hours after infection. Figure 1 is of cells without virus particles and figure 2 is of an infected cell. The nucleus and cytoplasm of the infected cells appear clearer than the cells without virus particles.

Studies on Water in the Hydration Chamber of a Modified Electron Microscope

1970 ◽  
Vol 28 ◽  
pp. 544-545
Author(s):  
R. C. Moretz ◽  
G. G. Hausner ◽  
D. F. Parsons
Keyword(s):  
Thin Film ◽  
Thin Films ◽  
Electron Microscope ◽  
Steady State ◽  
Room Temperature ◽  
Small Mass ◽  
Equilibrium Pressure ◽  
Biological Objects ◽  
Mechanical Pump ◽  
Differential Pumping

Use of the electron microscope to examine wet objects is possible due to the small mass thickness of the equilibrium pressure of water vapor at room temperature. Previous attempts to examine hydrated biological objects and water itself used a chamber consisting of two small apertures sealed by two thin films. Extensive work in our laboratory showed that such films have an 80% failure rate when wet. Using the principle of differential pumping of the microscope column, we can use open apertures in place of thin film windows.Fig. 1 shows the modified Siemens la specimen chamber with the connections to the water supply and the auxiliary pumping station. A mechanical pump is connected to the vapor supply via a 100μ aperture to maintain steady-state conditions.

Hydration Chamber for Jeolco 200 Kv Microscope

1970 ◽  
Vol 28 ◽  
pp. 542-543
Author(s):  
V. R. Matricardi ◽  
G. G. Hausner ◽  
D. F. Parsons
Keyword(s):  
Electron Microscope ◽  
Water Vapor ◽  
Vapor Pressure ◽  
Pressure Gradient ◽  
Room Temperature ◽  
List Type ◽  
Type Number ◽  
Equilibrium Vapor Pressure

In order to observe room temperature hydrated specimens in an electron microscope, the following conditions should be satisfied: The specimen should be surrounded by water vapor as close as possible to the equilibrium vapor pressure corresponding to the temperature of the specimen.The specimen grid should be inserted, focused and photo graphed in the shortest possible time in order to minimize dehydration.The full area of the specimen grid should be visible in order to minimize the number of changes of specimen required.There should be no pressure gradient across the grid so that specimens can be straddled across holes.Leakage of water vapor to the column should be minimized.

Ultrastructural comparison of prolactin adenomas of pituitary in men and women

1988 ◽  
Vol 46 ◽  
pp. 346-347
Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Pituitary Adenomas ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Men And Women ◽  
Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

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