Electron microscopy of the changes in the sperm head curvature of the palm squirrel (Funambulus Penanti)

Author(s):  
S. R. Bawa ◽  
R. Bawa ◽  
H. K. Bains

Examination of ultrathin sections of the spermatozoa recovered from the epididymis of the Indian palm squirrel (Funambulus penanti) indicates that the sperm head undergoes changes in its curvature during epididymal transit.Testis and epididymis of an adult male squirrel were dissected and small pieces of tissue fixed in 2.0% glutaraldehyde in 0.1 M phosphate buffer and post-fixed in osmium tetroxide. After dehydration in graded acetone the material was embedded in Araldite. Ultrathin sections were cut on a Reichert Jung Ultracut, picked-up on copper grids, stained with Reynold’s lead citrate-uranyl acetate and examined with a JEOL 1200 EX transmission electron microscope.Ultrathin sections of the caput epididymal spermatozoa reveal that their plasma membrane is adherent to the underlying acrosome (Figure 1). When these spermatozoa reach the corpus epididymis the plasma membrane surrounding the head becomes ruffled (Figure 2). The lifting-up of the plasma membrane around the head is restricted to the posterior bend of the acrosome.

Author(s):  
White Yvonne ◽  
Winslow Sheldon ◽  
James W. Townsend ◽  
Neil A. Littlefield

Spontaneous neoplasms rarely occur on the tails of BALB/cStCrlfC3H/Nctr mice, but the neoplasm most frequently observed is a locally invasive non-metastatic angiosarcoma. In this case a female mouse weighing 33.2 g, 706 days of age, presented a soft, red, irregularly-shaped mass, measuring 18 mm in its greatest dimension, in the subcutis of the base of the tail. A portion of the tail tumor was taken for electron microscopy and the remainder was processed for light microscopy. The tissue processed for electron microscopy was fixed in 4% cacodyl ate-buffered glutaraldehyde, post-fixed in 1% osmium tetroxide, dehydrated in a graded series of ethanol solutions, and embedded in Epon-Araldite resin mixture. Sections of 1 μm were stained with toluidine blue for light microscopy and ultrathin sections of 100 nm were stained with uranyl acetate and lead citrate, then examined with a Philips EM201 electron microscope .


Author(s):  
Masami Hokano ◽  
Tsunao Oh-I ◽  
Yoshie Narita ◽  
Hiroshi Sassa ◽  
Saburo Suzuki

Malacoplakia is a kind of granulomatous inflammation and characterized by the presence of calcium-stain positive granules (Michaelis-Gutmann bodies, hereinafter abbreviated as M-G bodies ) in the macrophages.In this report we want to say about the following articles:the ultrastruc tural findings in four cases of vesical malacoplakia;the ultrastructural morphogenesis of the M-G bodies;X-ray microanalytical studies which examine the change of the chemical component of M-G bodies, according to their developing stages; andacid phosphatase activity of lysosome within the malacoplakic macrophages.Biopsy materials taken from four cases with vesical malacoplakia were divided into 2 parts; one for a light microscopy and the other for an electron microscopy. The specimen for an electron microscopy was fixed in glutaraldehyde and osmium tetroxide, dehydrated in ethanol and then embedded in Epon 812. Ultrathin sections were doublestained with lead citrate and uranyl acetate and then subjected to routin TEM observation. For X-ray microanalysis was mounted an energy dispersive X-ray microanalyzer on a JEM-100 C type electron microscope. Ultrathin sections for microanalysis were cut 100-200nm thich and not stained.


Author(s):  
R.S. Demaree ◽  
L.J. Ackerman ◽  
D. L. Anderson

People who rapidly ascend to high terrestrial elevations may experience the “acute mountain sickness” syndrome. Speculation and limited evidence suggest that cerebral edema may play an important role in initiating and perpetuating this condition. We have recently demonstrated by electron microscopy that a mild cerebral edema develops in some Cebus apella monkeys rapidly transported to 14,110 feet. In the present study, Cebus apella monkeys were terminated at 1, 3, or 5 days after being shipped from sea level (160 feet) to 14,110 feet without acclimatization at intermediate altitudes.Thorotrast was administered IV 30 minutes prior to termination by perfusion or guillotine. Cerebral cortex was fixed by either perfusion or immersion in glutaraldehyde, and postfixed in osmium tetroxide. Following fixation, the tissues were dehydrated in ascending concentrations of ethanol followed by propylene oxide and embedded in Epon 812. Ultrathin sections were either not stained or doubly stained with uranyl acetate and lead citrate.


Author(s):  
J. C. Russ ◽  
E. McNatt

In order to study the retention of copper in cirrhotic liver, rats were made cirrhotic by carbon tetrachloride inhalation twice weekly for three months and fed 0.2% copper acetate ad libidum in drinking water for one month. The liver tissue was fixed in osmium, sectioned approximately 2000 Å thick, and stained with lead citrate. The section was examined in a scanning electron microscope (JEOLCO JSM-2) in the transmission electron mode.Figure 1 shows a typical area that includes a red blood cell in a sinusoid, a disse, and a portion of the cytoplasm of a hepatocyte which contains several mitochondria, peribiliary dense bodies, glycogen granules, and endoplasmic reticulum.


Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman

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.


Author(s):  
S.L. Asa ◽  
K. Kovacs ◽  
J. M. Bilbao ◽  
R. G. Josse ◽  
K. Kreines

Seven cases of lymphocytic hypophysitis in women have been reported previously in association with various degrees of hypopituitarism. We report two pregnant patients who presented with mass lesions of the sella turcica, clinically mimicking pituitary adenoma. However, pathologic examination revealed extensive infiltration of the anterior pituitary by lymphocytes and plasma cells with destruction of the gland. To our knowledge, the ultrastructural features of lymphocytic hypophysitis have not been studied so far.For transmission electron microscopy, tissue from surgical specimens was fixed in glutaraldehyde, postfixed in OsO4, dehydrated and embedded in epoxy-resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined with a Philips 300 electron microscope.Electron microscopy revealed adenohypophysial cells of all types exhibiting varying degrees of injury. In the areas of most dense inflammatory cell infiltration pituitary cells contained large lysosomal bodies fusing with secretory granules (Fig. 1), as well as increased numbers of swollen mitochondria, indicating oncocytic transformation (Fig. 2).


Author(s):  
Daniel C. Pease

It is reasonable to think that phospholipid micelles should be visible and identifiable in electron micrographs of ultrathin sections if only they can be preserved throughout the embedding process. The development of highly polar, water-containing, aminoplastic embedments has made this a likely possibility. With this in mind, an investigation of the lecithin-secreting, Type II pneumocytes of the lung is underway.Initially it has been easiest to recognize phospholipid micelles in lung tissue fixed first with glutaraldehyde, and then secondarily exposed to osmium tetroxide. However, the latter is not a necessary concomitant for micellar preservation. Conventional uranyl acetate and lead citrate staining is finally applied. Importantly, though, the micelles have been most easily seen in tissue embedded in 507. glutaraldehyde polymerized with urea, as described in detail by D.C. Pease and R.G. Peterson (J. Ultra- struct. Res., 41, 133, 1972). When oriented appropriately, the micellar units are seen as tiny, bilayer plates.


Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley

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.


Author(s):  
John H. L. Watson ◽  
Jessica Goodwin ◽  
E. Osborne Coates

Biopsies of lung were taken at operation from a patient with semi-acute diffuse pulmonary infiltrates for study by TEM and SEM. Tissue by light microscopy showed non-caseating granulomas consistent with sarcoidosis. Clinical evidence suggested a hypersensitivity reaction related to inhalation of substance of undetermined nature. Samples were fixed in glutaraldehyde, cacodylate-buffered. They were critical point dried and coated with Au-Pd for SEM, and were handled appropriately for TEM in Araldite. Sections were contrasted with uranyl acetate and lead citrate.


Author(s):  
F. A. Al-Bagdadi ◽  
C. W. Titkemeyer ◽  
J. E. Lovell

Skin biopsy samples were collected monthly from the lateral sides of 9 male Beagle dogs over a period of 1 year. The samples were fixed in 3% gluteral- dehyde and post fixed in 1% osmium tetroxide using phosphate or S-Collidine as buffers. They were dehydrated, embedded in Epon 812, sectioned with an LKB Ultrotome III, and stained with Reynolds' lead citrate and 1% uranyl acetate. They were examined and photographed by use of an RCA-3H electron microscope.The dermal papilla during anagen consisted of fibroblast-like cells some of which had cytoplasmic processes. The cytoplasm contained many mitochondria. (Fig. 1) The cells of the dermal papilla were either peripherally arranged spindle-shaped cells or polygonal cells. The basement membrane was either related to the fibroblast cytoplasmic processes or was free of the cytoplasmic processes but with a thick zone composed of fibers, granular ground substance and spaces.


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