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.

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.

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.

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 Comparative Study of Three Methods for the Ultrastructural Demonstration of Glycogen in Thin Sections

1971 ◽  
Vol 9 (3) ◽  
pp. 727-749
Author(s):  
M. V. VYE ◽  
D. A. FISCHMAN
Keyword(s):  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Uranyl Acetate ◽  
Sodium Chlorite ◽  
Lead Citrate ◽  
Thin Sections ◽  
Particle Form ◽  
Sequential Combination ◽  
Second Procedure ◽  
Glycogen Particles

In order to evaluate 3 staining methods for demonstration of glycogen in thin sections, 2 tissues containing an abundance of this carbohydrate in β-particle form were studied. Tissues were aldehyde-fixed, postfixed in osmium tetroxide, embedded in Araldite and sectioned in the usual manner without special precautions. The first method for staining thin sections employed a sequential combination of periodic acid, thiosemicarbazide and osmium tetroxide vapour, while in the second procedure a silver protein solution was substituted for the osmium tetroxide vapour. The third technique utilized periodic acid, sodium chlorite and uranyl acetate, also in sequential combination. Each method yielded glycogen particles of greater electron density than were seen in sections stained by the usual uranyl acetate-lead citrate procedure. Under high magnification, considerable method-dependent variation in the appearance of the glycogen granules was noted. Particulate substructure, only faintly visible in routinely stained sections, was easily resolved with the periodic acid-thiosemicarbazide-silver protein technique. Conversely, periodic acid-thiosemicarbazide-osmium tetroxide completely obscured this substructure. With periodic acid-sodium chlorite-uranyl acetate, glycogen particles appeared larger, more confluent, and of a less regular outline than with the other methods. Sections were also stained by incubation in periodic acid prior to treatment with lead citrate. The alteration in appearance of the glycogen granules produced by this modification was so great that high-resolution analysis of particle size and substructure could not be undertaken. The usefulness of the procedures investigated here resides in their ability to stain glycogen in thin sections in an intense and selective manner.

scholarly journals AN ELECTRON MICROSCOPIC STUDY OF NUCLEAR ELIMINATION FROM THE LATE ERYTHROBLAST

1967 ◽  
Vol 33 (3) ◽  
pp. 625-635 ◽  
Author(s):  
Ehud Skutelsky ◽  
David Danon
Keyword(s):  
Bone Marrow ◽  
Osmium Tetroxide ◽  
Mitotic Division ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Whole Process ◽  
Narrow Passage ◽  
Rounded Form

The process of expulsion of the nucleus during the transformation of the late erythroblast to reticulocyte is described. Erythroid clones taken from the spleen of lethally irradiated mice transplanted with syngeneic bone marrow were used. 10–12-day old isolated clones were fixed in glutaraldehyde, then in osmium tetroxide. Ultra-thin sections were stained with uranyl acetate and/or lead citrate before examination. Late (orthochromatic) erythroblasts develop pseudopod-like cytoplasmic protrusions into one of which the nucleus gradually penetrates, being deformed by the extrusion through the relatively narrow passage. During the whole process, mitochondria and vesicular and membranous elements are concentrated in the cytoplasm. Once outside the cell, the nucleus reassumes its rounded form. It is surrounded by a narrow rim of cytoplasm and structurally altered plasma membrane and is connected to the rest of the cell by a bridge. Elongated vacuoles appear within this bridge, with a resulting release of the enveloped nucleus which is soon phagocytized by macrophages; this leaves behind the newly formed reticulocyte. During this process, the cytoplasmic protrusions, the agglomeration of mitochondria, and the mode of separation of the nucleus from the rest of the cell are similar to those occurring in mitotic division.

Unusual Intranuclear Structures in Chick Sympathetic Neurons

1967 ◽  
Vol 25 ◽  
pp. 188-189
Author(s):  
E. B. Masurovsky ◽  
H. H. Benitez ◽  
M. R. Murray
Keyword(s):  
Electron Microscope ◽  
Sympathetic Neurons ◽  
Uranyl Acetate ◽  
Sympathetic Ganglia ◽  
Lead Citrate ◽  
Thin Sections ◽  
Cns Neurons ◽  
Mammalian Cns

Recent light- and electron microscope studies concerned with the effects of D2O on the development of chick sympathetic ganglia in long-term, organized culture revealed the presence of rod-like fibrillar formations, and associated granulofibrillar bodies, in the nuclei of control and deuterated neurons. Similar fibrillar formations have been reported in the nuclei of certain mammalian CNS neurons; however, related granulofibrillar bodies have not been previously described. Both kinds of intranuclear structures are observed in cultures fixed either in veronal acetate-buffered 2%OsO4 (pH 7. 4), or in 3.5% glutaraldehyde followed by post-osmication. Thin sections from such Epon-embedded cultures were stained with ethanolic uranyl acetate and basic lead citrate for viewing in the electron microscope.

Fine Structure of Interdental Cells in the Mouse Cochlea

1970 ◽  
Vol 28 ◽  
pp. 140-141
Author(s):  
Roberta M. Bruck
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Young Adult ◽  
Uranyl Acetate ◽  
Ground Substance ◽  
Tectorial Membrane ◽  
Lead Citrate ◽  
Unusual Structure ◽  
Thin Sections ◽  
Collagenous Fibers

An unusual structure in the cochlea is the spiral limbus; this periosteal tissue consists of stellate fibroblasts and collagenous fibers embedded in a translucent ground substance. The collagenous fibers are arranged in vertical columns (the auditory teeth of Haschke). Between the auditory teeth are interdental furrows in which the interdental cells are situated. These epithelial cells supposedly secrete the tectorial membrane.The fine structure of interdental cells in the rat was reported by Iurato (1962). Since the mouse appears to be different, a description of the fine structure of mouse interdental cells' is presented. Young adult C57BL/6J mice were perfused intervascularly with 1% paraformaldehyde/ 1.25% glutaraldehyde in .1M phosphate buffer (pH7.2-7.4). Intact cochlea were decalcified in .1M EDTA by the method of Baird (1967), postosmicated, dehydrated, and embedded in Araldite. Thin sections stained with uranyl acetate and lead citrate were examined in a Phillips EM-200 electron microscope.

Ultrastructure of myoepithelial cell in parotid gland

1988 ◽  
Vol 46 ◽  
pp. 342-343
Author(s):  
C. N. Sun
Keyword(s):  
Parotid Gland ◽  
Osmium Tetroxide ◽  
Individual Cell ◽  
Branching Processes ◽  
Muscle Cells ◽  
Myoepithelial Cells ◽  
Uranyl Acetate ◽  
Thin Sections ◽  
Gland Carcinoma ◽  
Parotid Gland Carcinoma

Myoepithelial cells have been observed in the prostate, harderian, apocrine, exocrine sweat and mammary glands. Such cells and their numerous branching processes form basket-like structures around the glandular acini. Their shapes are quite different from structures seen either in spindleshaped smooth muscle cells or skeletal muscle cells. These myoepithelial cells lie on the epithelial side of the basement membrane in the glands. This presentation describes the ultrastructure of such myoepithelial cells which have been found also in the parotid gland carcinoma from a 45-year old patient.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4 percent glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1 percent buffered osmium tetroxide for 1 hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate. Ultrastructurally, the pattern of each individual cell showed wide variations.

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