Intracellular and Extracellular Acid Hydrolase Activity in the Drosophila Melanogaster Ovary

1982 ◽  
Vol 40 ◽  
pp. 242-243
Author(s):  
Lawrence G. Altman ◽  
R. Witkus ◽  
G. M. Vernon
Keyword(s):  
Drosophila Melanogaster ◽  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Acid Hydrolase ◽  
Lead Citrate ◽  
Wild Type ◽  
Thin Sections

Ovaries from wild type Drosophila melanogaster were incubated for acid phosphatase activity (pH 5). Following fixation in either 3.0% or 6.0% glutaraldehyde prepared in 0.1M cacodyiate buffer. Post-fixation was done in 1.0% osmium tetroxide and tissues were subjected to a graded series of acetone and infiltrated with Epon-Araldite. Control tissues were incubated without the substrate, in this case Bglycerophosphate. Ultrastructural integrity was checked against tissue prepared by omitting the incubation period completely. Thin and semi-thin sections were cut on an LKB ultramicrotome. In some instances, observations were recorded without uranyl acetate and lead citrate counterstaining.

scholarly journals LOCALIZATION OF ACID PHOSPHATASE ACTIVITY IN HEPATIC LYSOSOMES BY MEANS OF ELECTRON MICROSCOPY

1961 ◽  
Vol 9 (4) ◽  
pp. 773-784 ◽  
Author(s):  
E. Essner ◽  
Alex B. Novikoff
Keyword(s):  
Electron Microscopy ◽  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Human Liver ◽  
Osmium Tetroxide ◽  
Frozen Sections ◽  
Fixed Tissue ◽  
Thin Sections ◽  
Lipofuscin Granules

Samples of liver from untreated rats, from rats infused with unconjugated bilirubin, and from biopsies of human liver were fixed overnight in cold formol-calcium. Frozen sections were stained for acid phosphatase activity by the Gomori lead-glycerophosphate procedure. Small blocks of fixed tissue were also incubated in this medium. These were then treated briefly with osmium tetroxide, dehydrated, and embedded in methacrylate. Thin sections were studied by electron microscopy. The sites of reaction product of acid phosphatase activity as visualized in electron micrographs are consistent with those seen in frozen sections studied by light microscopy. They indicate that the pericanalicular bodies of parenchymatous cells, the large spherical bodies of Kupffer cells, the microbodies appearing after bilirubin infusion and lipofuscin granules belong to the class of cytoplasmic organelles called lysosomes by de Duve.

Membranous alterations in the cytoplasm and nucleus in a primitive neuroectodermal tumor of the brain

1978 ◽  
Vol 36 (2) ◽  
pp. 628-629
Author(s):  
C. N. Sun ◽  
C. Araoz ◽  
H. J. White
Keyword(s):  
Nuclear Envelope ◽  
Tumor Cells ◽  
Osmium Tetroxide ◽  
Primitive Neuroectodermal Tumor ◽  
Uranyl Acetate ◽  
Neuroectodermal Tumor ◽  
Annulate Lamellae ◽  
Lead Citrate ◽  
Thin Sections ◽  
The Brain

The ultrastructure of a cerebral primitive neuroectodermal tumor has been reported previously. In the present case, we will present some unusual previously unreported membranous structures and alterations in the cytoplasm and nucleus of the tumor cells.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1% buffered osmium tetroxide for one hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate.In the cytoplasm of the tumor cells, we found paired cisternae (Fig. 1) and annulate lamellae (Fig. 2) noting that the annulate lamellae were sometimes associated with the outer nuclear envelope (Fig. 3). These membranous structures have been reported in other tumor cells. In our case, mitochondrial to nuclear envelope fusions were often noted (Fig. 4). Although this phenomenon was reported in an oncocytoma, their frequency in the present study is quite striking.

An Electron Microscopic Investigation of the Pathogenesis of Hemorrhage Induced by Rattlesnake Venom

1974 ◽  
Vol 32 ◽  
pp. 56-57
Author(s):  
Charlotte L. Ownby ◽  
Robert A. Kainer ◽  
Anthony T. Tu
Keyword(s):  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Rattlesnake Venom ◽  
Diamondback Rattlesnake

One of the significant changes induced by the injection of rattlesnake (Crotalidae) venom is hemorrhage. Since crotaline antivenin does not prevent such local tissue damage, a more effective treatment of snakebite is needed. To aid in the development of such a treatment the pathogenesis of venom-induced hemorrhae was investigated.Swiss-Webster white mice were injected intramuscularly with Western diamondback rattlesnake (Crotalus atrox) venom. Two minutes after the injection, muscle tissue was obtained by bioosy from the thigh and fixed in 6% glutaraldehyde in Milloniq's phosphate buffer (DH 7.4, 2 hrs., 4°C). After post-fixation in 2% osmium tetroxide in Milloniq's phosphate buffer (pH 7.4, 1hr., 4°C) the tissue was dehydrated routinely in ethanol and embedded in Epon 812. The thin sections were stained with uranyl acetate in methanol and lead citrate then observed with either a Zeiss EM 9A or an Hitachi HS-8 electron microscope.

Intracisternal microtubules in proximal tubule cells of duck kidney: A serendipitous finding

1987 ◽  
Vol 45 ◽  
pp. 898-899
Author(s):  
M.K. Bhatnagar ◽  
S.M. Snelgrove-Hobson ◽  
P.V.V. Prasada Rao
Keyword(s):  
Electron Microscope ◽  
Proximal Tubule ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Kidney Cortex ◽  
Lead Citrate ◽  
Cell Organelles ◽  
Thin Sections ◽  
Proximal Tubule Cells ◽  
Tubule Cells

Cytpplasmic microtubules, ubiquitous cell organelles, lie free in the cytosol without being compartmented off by membranes. In rare instances, however, intracisternal microtubules (ICM) have been reported in ganglionic neurons of aging dogs and in the proximal tubule cells (PTC) of rabbit kidneys. We report here the presence of ICM in the PTC of the kidneys of Peking ducks (Anas platyrhynhos).A total of 106 Peking ducks (24 males and 24 females of 9 months, 48 females of 15 months, 6 males of 30 months and 2 males and 2 females of 48 months of age) were anesthetized and exsanguinated. Pieces of kidney cortex were fixed in 4% glutaraldehyde in 0.5M phosphate buffer (pH 7.3) at 29°C for four hours. Samples were post fixed in 2% osmium tetroxide for two hours. One micron sections of Epon-embedded cortices were stained with toluidine blue and thin sections (70-70 nm) contrasted with uranyl acetate and lead citrate were examined with a JEOL 100-S electron microscope at 80 kV.

Neural Degeneration in the Spiral Ganglia of C57BL/6 Mice

1988 ◽  
Vol 46 ◽  
pp. 136-137
Author(s):  
Glenn M. Cohen ◽  
Joseph S. Grasso
Keyword(s):  
Osmium Tetroxide ◽  
Initial Study ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Thin Sections ◽  
Neural Degeneration ◽  
Age Related ◽  
Sensory Hair Cell ◽  
Spiral Ganglia ◽  
Old Mice

C57BL/6 mice, along with several other mouse genotypes, have served as models for human presbycusis (age-related hearing losses). C57BL/6 mice and their genetic substrain C57/bl6 show progressively severe hearing losses, starting as early as 30 days postnatally. The hearing losses result from sweeping sensory (hair cell) and neural degeneration that begins in the basal end and advances apically. For the initial study of the spiral ganglia C57BL/6 mice, our two objectives were to develop criteria for identifying the different degenerative stages and determine Whether the same degenerative stages repeat themselves in both young and old mice.Female C57BL/6 mice of seven ages, ranging from 1 1/2 to 32 months, were examined. The ears were exposed to the fixative (3.5% glutaraldehyde [GA]) within cne min after sacrifice. The cochleae were decalcified in 5% EDTA in 3.5% GA, postfixed in 1% osmium tetroxide, and embedded in Araldite 502 epoxy. Semithin (1μ) sections were stained with toluidine blue or p-phenylenediamine; thin sections were stained with uranyl acetate and lead citrate.

Cystoid Degeneration of Mouse Pituitary – Ultrastructural Study

1980 ◽  
Vol 38 ◽  
pp. 462-463
Author(s):  
Annette M. Andrews ◽  
Alex M. Cameron ◽  
James W. Townsend ◽  
Winslow G. Sheldon
Keyword(s):  
Electron Microscope ◽  
Toluidine Blue ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Pars Intermedia ◽  
Lead Citrate ◽  
Thin Sections ◽  
Resin Mixture ◽  
Mouse Pituitary ◽  
Two Sides

Pituitaries of 6 C3H/HEJ MTV+ mice about 660 days of age were fixed by immersion in cacodylate-buffered 4% glutaraldehyde, post-fixed in 1% osmium tetroxide, stained en block with aqueous uranyl acetate, dehydrated in a graded series of ethanol solutions, cleared in acetone, and embedded in an Epon-Araldite resin mixture. Semi-thin (1 μm) sections were taken of the entire face of the block, and stained with toluidine blue. Subsequently, thin sections (100 nm) were prepared from mesas of the two sides of the pars distal is and one mesa of the pars intermedia. Thin sections were stained with ethanolic uranyl acetate followed by Sato's lead citrate (1), then examined on a Philips EM201 electron microscope.

A New Smaller Sized Virus-Like Particle in Drosophila Melanogaster

2001 ◽  
Vol 7 (S2) ◽  
pp. 742-743
Author(s):  
Jeffrey G. Ault ◽  
Ellen Shimakawa
Keyword(s):  
Drosophila Melanogaster ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Freeze Substitution ◽  
Uranyl Acetate ◽  
High Pressure Freezing ◽  
Chromosome Behavior ◽  
Thin Sections ◽  
Behavior Study ◽  
Virus Like Particle

During a chromosome behavior study involving high-pressure freezing (HPF)/freeze substitution (FS) of Drosophila melanogaster testes, we discovered quasi-crystalline inclusions in the nuclei of adjacent gut epithelial cells (Fig. 1). The HPF and FS protocols were standard. The viscera of adult flies were packed in yeast paste for HPF. The tissue was fixed by FS with 1% osmium tetroxide in acetone for 72 hours at -90° C then 48 hours at -60° C. Afterwards, it was washed several times at room temperature in 100% acetone and embedded in Epon/Araldite. Thin sections were cut and stained with uranyl acetate and lead. As expected with HPF/FS, the material was well-preserved with straight microtubules, smooth membranes, dense mitochondria, and abundant ribosomes both on the rough endoplasmic reticulum and in the full cytoplasm (Fig. 1).The inclusions consisted of virus-like particles packed loosely together in orderly arrays. Particles were usually hexagonally packed with spaces disrupting the periodicity (Figs. 2 and 3).

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