On the variability of spherosome-like bodies in Phaseolus vulgaris

1970 ◽  
Vol 48 (8) ◽  
pp. 1477-1480 ◽  
Author(s):  
A. K. Mishra ◽  
J. Ross Colvin
Keyword(s):  
Electron Microscope ◽  
Phaseolus Vulgaris ◽  
Electron Density ◽  
Potassium Permanganate ◽  
Osmium Tetroxide ◽  
Marked Variation ◽  
Phaseolus Vulgaris L ◽  
Shoot Apices ◽  
Distribution Shape ◽  
Physiological Heterogeneity

Root and shoot apices and primary leaves of 3- to 4-day-old seedlings of Phaseolus vulgaris L. were examined in the electron microscope after fixation with (a) glutaraldehyde followed by osmium tetroxide, (b) glutaraldehyde followed by potassium permanganate, (c) osmium tetroxide alone, and (d) potassium permanganate alone. The distribution, shape, and substructure of the spherosome-like bodies in the cytoplasm varied markedly depending upon the tissue, its age, and the fixation used. Shapes varied from stellate through polygonal to spherical. The electron density of the interior of the organelles varied from uniformly dark after osmium tetroxide fixation to almost none after permanganate fixation. In addition to differences caused by fixatives, marked variation of the electron density of the same organelles with identical fixation was observed. Structural, and possibly physiological, heterogeneity seems to be characteristic of spherosomes but in spite of their variability, they may be recognized in the electron microscope as a group of similar structures distinct from other organelles.

scholarly journals The ultracytochemical demonstration of a formaldehyde-resistant dehydrogenase of leuco nitroxyl analogues.

1975 ◽  
Vol 23 (11) ◽  
pp. 840-854 ◽  
Author(s):  
W A Shannon ◽  
H L Wasserkrug ◽  
R E Plapinger ◽  
D Lynm ◽  
A M Seligman
Keyword(s):  
Skeletal Muscle ◽  
Electron Microscope ◽  
Dehydrogenase Activity ◽  
Electron Density ◽  
Osmium Tetroxide ◽  
Tetrazolium Salt ◽  
Hydrogen Ions ◽  
Tetrazolium Chloride

A dehydrogenase which is relatively stable in formaldehyde fixative is demonstrated ultracytochemically by the reduction of various leuco nitroxyl analogues in rat hepatic, renal, myocardial, skeletal muscle and prostatic tubuloalveolar glandular tissues. The nonosmiophilic tetrazolium salt, t-(2'-benzothiazolyl)-5-styryl-3-(4'-phtalhydrazidyl) tetrazolium chloride, is subsequently reduced to an insoluble osmiophilic formazan by the hydrogen ions resulting from the dehydrogenase activity. Exposure of the formazan to osmium tetroxide results in electron density enabling visualization of the reaction product in the electron microscope. Known inhibitors of various dehydrogenases were utilized in an attempt to determine the existence and/or extent of any specific characteristics of the dehydrogenase(s) involved.

scholarly journals ON THE FINE STRUCTURE AND COMPOSITION OF THE NUCLEAR ENVELOPE

1961 ◽  
Vol 11 (3) ◽  
pp. 559-570 ◽  
Author(s):  
R. W. Merriam
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Nuclear Envelope ◽  
Pore Structure ◽  
Potassium Permanganate ◽  
Osmium Tetroxide ◽  
Rana Pipiens ◽  
Total Density ◽  
Thin Sections ◽  
Whole Mounts

Nuclei from nearly ripe eggs of Rana pipiens were isolated and cleaned in 0.1 M KCl. The whole nucleus was then digested to various degrees with ribonuclease or trypsin, followed by washing and fixation in either osmium tetroxide or potassium permanganate. The nuclear envelope was dissected off, placed on a grid, air dried, and compared with undigested controls in the electron microscope. Some envelopes were dehydrated, embedded in methacrylate, and sectioned. Annuli around "pores" are composed of a substance or substances, at least partially fibrillar, which is preserved by osmium but lost during permanganate fixation. Material within the "pores" is also preserved by osmium but partially lost after permanganate. No evidence of granules or tubules in the annuli was found in air dried mounts although a granular appearance could be seen in tangentially oriented thin sections. Thin sections of isolated envelopes give evidence of diffuse material within the "pores" as well as a more condensed diaphragm across their waists. In whole mounts of the envelope the total density within "pores" is relatively constant from "pore" to "pore." All material within "pores," including the condensed diaphragm, is removable by trypsin digestion. Wispy material from the "pore" structure projects into the nucleus and annular material extends into the cytoplasm. Both annular and diaphragm materials remain with the envelope when it is isolated and are thus considered a part of its structure, not merely evidences of material passing through. There is no evidence of ribonuclease-removable material in any part of the "pore" complex.

scholarly journals AN ATYPICAL CRISTA RESEMBLING A "TIGHT JUNCTION" IN BEAN ROOT MITOCHONDRIA

1968 ◽  
Vol 39 (1) ◽  
pp. 35-42 ◽  
Author(s):  
Eldon H. Newcomb ◽  
Martin W. Steer ◽  
Peter K. Hepler ◽  
William P. Wergin
Keyword(s):  
Phaseolus Vulgaris ◽  
Tight Junction ◽  
Tight Junctions ◽  
Osmium Tetroxide ◽  
Root Tip ◽  
Mitochondrial Matrix ◽  
Phaseolus Vulgaris L ◽  
Animal Cells ◽  
Tip Cells ◽  
Root Tip Cells

The conformation and structure of an atypical crista found in a small percentage of the mitochondria in root tip cells of Phaseolus vulgaris L. have been studied electron microscopically in material fixed in glutaraldehyde followed by osmium tetroxide. In its transformation into an atypical crista, a normal crista elongates, broadens, and flattens, and the inner leaflets of its apposed unit membranes appear to fuse in a manner analogous to the formation of "tight junctions" between certain animal cells. The result is a large platelike, quintuple-layered structure, 240–260 A thick, whose long axis parallels that of the mitochondrion. The outer layers of the "plate," bordering on the mitochondrial matrix, are thickened and exhibit striking patterns in the micrographs. The structure of the plate is compared with that previously described for tight junctions between animal cells.

scholarly journals Electron Microscope Studies of the Human Epidermis The Clear Cell of Masson (Dendritic Cell or Melanocyte)

1958 ◽  
Vol 4 (6) ◽  
pp. 679-684 ◽  
Author(s):  
Wallace H. Clark ◽  
Richard G. Hibbs
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Dendritic Cell ◽  
Potassium Permanganate ◽  
Osmium Tetroxide ◽  
Clear Cell ◽  
Human Epidermis ◽  
Intercellular Bridges ◽  
Distinct Cell

The human epidermis has been studied by electron microscopy following osmium tetroxide and potassium permanganate fixation. An anatomically distinct cell in the human epidermis has been demonstrated with features similar to the melanocyte of the hair bulb described by Barnicot, Birbeck and Cuckow (3). It is dendritic in form and does not contain tonofilaments. "Intercellular bridges" are not formed. The mitochondria are larger and more numerous than those of other epidermal cells and the endoplasmic reticulum is more complex. Some of these cells contain melanin but others are melanin-free. The cell has been interpreted as being identical with the dopa-positive, clear cell of Masson (dendritic cell of Bloch or melanocyte). We have found that many membranous structures in the human epidermis are better preserved by permanganate fixation than by osmium tetroxide fixation.

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.

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.

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.

Ultrastructure of Testicular Spermatozoon of the Fish Oreochromis Niloticus

1988 ◽  
Vol 46 ◽  
pp. 278-279
Author(s):  
T. Guha ◽  
A. Q. Siddiqui ◽  
P. F. Prentis
Keyword(s):  
Electron Microscope ◽  
Oreochromis Niloticus ◽  
Osmium Tetroxide ◽  
Sperm Cells ◽  
Adult Fish ◽  
Testicular Sperm ◽  
Thin Sections ◽  
Important Fish ◽  
Testicular Spermatozoon ◽  
Diamond Knife

Tilapia, Oreochromis niloticus, is an economically important fish in Saudi Arabia. Elucidation of reproductive biology of this species is necessary for successful breeding program. In this paper we describe fine structure of testicular sperm cells in O, niloticus.Testes from young adult fish were fixed in gluteraldehyde (2%) and osmium tetroxide (1%), both in cacodyl ate buffer. Specimens were processed in the conventional way for electron microscopy and thin sections of tissues (obtained by cutting the blocks with a diamond knife) were stained by ura- nyl acetate and lead citrate. These were examined in a Carl Zeiss electron microscope operated at 40 kV to 60 kV. Sperm cells were obtained from testes by squeezing them in cacodyl ate buffer. They were fixed in gluteraldehyde (2%) in the same buffer, air dried, gold coated and then examined in a Philips scanning electron microscope (SEM) operated at 25kV.The spermatozoon of O. niloticus is consisting of head, midpiece and tail (Fig. 1).

Selective Staining of Acid Mucopolysaccharides By Ruthenium Red

1968 ◽  
Vol 26 ◽  
pp. 38-39
Author(s):  
J. H. Luft
Keyword(s):  
Electron Microscope ◽  
Light Microscopy ◽  
Light Microscope ◽  
Osmium Tetroxide ◽  
Single Cells ◽  
Ruthenium Red ◽  
Collagen Fibers ◽  
Selective Staining ◽  
Pectic Substances ◽  
Solid Tissues

Ruthenium red is one of the few completely inorganic dyes used to stain tissues for light microscopy. This novelty is enhanced by ignorance regarding its staining mechanism. However, its continued usefulness in botany for demonstrating pectic substances attests to selectivity of some sort. Whether understood or not, histochemists continue to be grateful for small favors.Ruthenium red can also be used with the electron microscope. If single cells are exposed to ruthenium red solution, sufficient mass can be bound to produce observable density in the electron microscope. Generally, this effect is not useful with solid tissues because the contrast is wasted on the damaged cells at the block surface, with little dye diffusing more than 25-50 μ into the interior. Although these traces of ruthenium red which penetrate between and around cells are visible in the light microscope, they produce negligible contrast in the electron microscope. However, its presence can be amplified by a reaction with osmium tetroxide, probably catalytically, to be easily visible by EM. Now the density is clearly seen to be extracellular and closely associated with collagen fibers (Fig. 1).

Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

1968 ◽  
Vol 26 ◽  
pp. 20-21
Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher
Keyword(s):  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Undifferentiated Carcinoma ◽  
Uranyl Acetate ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
North West ◽  
X Irradiation ◽  
Tissue Specimens

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.

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