scholarly journals CYTOCHEMISTRY AND ELECTRON MICROSCOPY

1963 ◽  
Vol 17 (1) ◽  
pp. 19-58 ◽  
Author(s):  
David D. Sabatini ◽  
Klaus Bensch ◽  
Russell J. Barrnett
Keyword(s):  
Electron Microscopy ◽  
Alkaline Phosphatase ◽  
Fine Structure ◽  
Acid Phosphatase ◽  
Adenosine Triphosphatase ◽  
Osmium Tetroxide ◽  
Enzyme Histochemistry ◽  
Succinic Dehydrogenase ◽  
Optimal Conditions ◽  
Fixation Procedure

The aldehydes introduced in this paper and the more appropriate concentrations for their general use as fixatives are: 4 to 6.5 per cent glutaraldehyde, 4 per cent glyoxal, 12.5 per cent hydroxyadipaldehyde, 10 per cent crotonaldehyde, 5 per cent pyruvic aldehyde, 10 per cent acetaldehyde, and 5 per cent methacrolein. These were prepared as cacodylate- or phosphate-buffered solutions (0.1 to 0.2 M, pH 6.5 to 7.6) that, with the exception of glutaraldehyde, contained sucrose (0.22 to 0.55 M). After fixation of from 0.5 hour to 24 hours, the blocks were stored in cold (4°C) buffer (0.1 M) plus sucrose (0.22 M). This material was used for enzyme histochemistry, for electron microscopy (both with and without a second fixation with 1 or 2 per cent osmium tetroxide) after Epon embedding, and for the combination of the two techniques. After fixation in aldehyde, membranous differentiations of the cell were not apparent and the nuclear structure differed from that commonly observed with osmium tetroxide. A postfixation in osmium tetroxide, even after long periods of storage, developed an image that—notable in the case of glutaraldehyde—was largely indistinguishable from that of tissues fixed under optimal conditions with osmium tetroxide alone. Aliesterase, acetylcholinesterase, alkaline phosphatase, acid phosphatase, 5-nucleotidase, adenosine triphosphatase, and DPNH and TPNH diaphorase activities were demonstrable histochemically after most of the fixatives. Cytochrome oxidase, succinic dehydrogenase, and glucose-6-phosphatase were retained after hydroxyaldipaldehyde and, to a lesser extent, after glyoxal fixation. The final product of the activity of several of the above-mentioned enzymes was localized in relation to the fine structure. For this purpose the double fixation procedure was used, selecting in each case the appropriate aldehyde.

A Histological and Histochemical Study of the Brown and Yellow Adipose Tissue of the Bat, Hipposideros speoris

1959 ◽  
Vol s3-100 (51) ◽  
pp. 369-375
Author(s):  
J. C. GEORGE ◽  
J. EAPEN
Keyword(s):  
Alkaline Phosphatase ◽  
Adipose Tissue ◽  
Acid Phosphatase ◽  
Adenosine Triphosphatase ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Histological Structure ◽  
Brown Adipose ◽  
Aldehydes And Ketones ◽  
Sulphydryl Groups

A study of the histology and histochemical reactions for lipase, alkaline phosphatase, acid phosphatase, adenosine triphosphatase, succinic dehydrogenase, lactic dehydrogenase, phospholipids, cholesterol, sulphydryl groups, and water-insoluble aldehydes and ketones in the brown and yellow adipose tissue of the bat (Hipposideros speoris) revealed that the two types of adipose tissue differ in histological structure as well as physiological activity. The histological structure of the two types of adipose tissue was found to be different, resembling that of the two corresponding types of the rat. The brown adipose tissue showed a higher concentration of succinic dehydrogenase, lactic dehydrogenase, phospholipids, cholesterol, and sulphydryl groups. No detectable difference between brown and yellow adipose tissue was, however, found with respect to lipase, alkaline phosphatase, acid phosphatase, adenosine triphosphatase, and water-insoluble aldehydes and ketones.

scholarly journals Enzyme histochemistry of developing rat oral mucosa.

1981 ◽  
Vol 29 (1) ◽  
pp. 57-64 ◽  
Author(s):  
S Sjögren ◽  
L Hammarström ◽  
A Larsson
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Lactate Dehydrogenase ◽  
Succinate Dehydrogenase ◽  
Oral Mucosa ◽  
Adenosine Triphosphatase ◽  
Enzyme Histochemistry ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Developing Rat

The oral mucosa of developing and mature rats was analyzed histochemically for regional enzyme differences. The following enzymes were studied: nonspecific alkaline phosphatase (alkpase), acid phosphatase (acidpase), 5'-nucleotidase (AMPase), adenosine triphosphatase (ATPase), succinate dehydrogenase (SDH), lactate dehydrogenase (LDH), and glucose-6-phosphate dehydrogenase (G-6-pDH). All enzymes were active in the oral mucosa, but regional as well as tissue variations were observed. Epithelium in all regions showed acidpase staining. Oxidoreductases were found in all regions with variations within the epithelium. The epithelium of specific regions stained for alkpase and AMPase, while adjacent epithelium did not. We suggest that the alkpase and AMPase activities are associated with specific functions of the epithelium in these regions.

scholarly journals The fine structure produced in cells by primary fixatives 2. Potassium dichromate

1965 ◽  
Vol s3-106 (73) ◽  
pp. 15-21
Author(s):  
JOHN R. BAKER
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Fine Structure ◽  
Golgi Apparatus ◽  
Nuclear Membrane ◽  
Osmium Tetroxide ◽  
Potassium Dichromate ◽  
Potassium Dichromate Solution ◽  
Zymogen Granules ◽  
Mouse Pancreas

The exocrine cells of the mouse pancreas were fixed in potassium dichromate solution, embedded in araldite or other suitable medium, and examined by electron microscopy. Almost every part of these cells is seriously distorted or destroyed by this fixative. The ergastoplasm is generally unrecognizable, the mitochondria and zymogen granules are seldom visible, and no sign of the plasma membrane, microvilli, or Golgi apparatus is seen. The contents of the nucleus are profoundly rearranged. It is seen to contain a large, dark, irregularly shaped, finely granular object; the evidence suggests that this consists of coagulated histone. The sole constituent of the cell that is well fixed is the inner nuclear membrane. The destructive properties of potassium dichromate are much mitigated when it is mixed in suitable proportions with osmium tetroxide or formaldehyde.

Cytological and cytochemical studies on Cytamoeba bacterifera Labbé, 1894

Parasitology ◽  
1964 ◽  
Vol 54 (1) ◽  
pp. 121-124 ◽  
Author(s):  
D. L. Lehmann
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Toluidine Blue ◽  
Osmium Tetroxide ◽  
Central Portion ◽  
Central Mass ◽  
Living Entity ◽  
Peripheral Ring ◽  
American Foundation ◽  
Long Rod

Cytamoeba bacterifera from urodeles (Batrachoseps attenuatus, Aneides lugubris, A. flavipunctatus and Dicamptodon ensatus) show a thin peripheral ring of DNA and a less definite, heterogeneous central portion after Feulgen treatment; toluidine blue reveals a similar configuration. Exposure to DNA-ase and RNA-ase, followed by toluidine blue and Feulgen treatment, substantiates the localization of DNA; RNA is present in small quantity and restricted to the periphery of the parasite.Mitochondria (2–5μ long rod-shaped, central bodies) can be detected with Janus Green B, and osmium tetroxide yields a dark central mass of rods and granules as well as peripheral granules; Sudan Black IV is without action. Zymohexase and acid phosphatase can be detected by cytochemical means, but peroxidase, lipase, urease and alkaline phosphatase were not noted.From the evidence presented, C. bacterifera is considered a living entity and is tentatively relegated to the Piroplasmida.The writer would like to express his appreciation to Dr R. S. Bray of the American Foundation for Tropical Medicine, Harbel, Liberia, for his counsel and suggestions.

scholarly journals Retrieval of cryostat section for comparison of histochemistry and quantitative electron microscopy in a muscle fiber.

1977 ◽  
Vol 25 (10) ◽  
pp. 1169-1177 ◽  
Author(s):  
B R Eisenberg ◽  
A M Kuda
Keyword(s):  
Electron Microscopy ◽  
Muscle Fiber ◽  
Cross Sections ◽  
Adenosine Triphosphatase ◽  
Volume Fraction ◽  
Cryostat Section ◽  
Succinic Dehydrogenase ◽  
Medial Gastrocnemius ◽  
Dehydrogenase Reaction ◽  
Mitochondrial Volume

A method is presented that can be used to perform histochemical and morphometric analyses on the same muscle fiber. Freshly dissected fibers from medial gastrocnemius muscle of adult guinea pig were kept at a resting length and rapidly frozen. Serial frozen cross-sections were cut and reacted for myofibrillar adenosine triphosphatase and succinic dehydrogenase. The adjacent section, while still frozen, was immersed into 20 degrees C glutaraldehyde fixative to which EGTA was added to minimize artifactious contraction. The fixed section was processed for electron microscopy and the section rotated before thin sectioning to give longitudinal sections enabling study of sarcomeres. Ultrastructure was well-preserved despite slight disorganization of the contractile filaments and some vesiculation of the sarcoplasmic reticulum. The Z line width was measured and the mitochondrial volume fraction estimated by point counting morphometry from 89 fibers. The fibers with dark myofibrillar adenosine triphosphatase staining have Z widths of 547 +/- 165 A (n=69) and thoshosphatase staining have Z widths of 547 +/- 165 A (n=69) and those with light stain have 1023 +/- 113 A (n=20). The density of the succinic dehydrogenase reaction product in the fibers was divided into dark and light and the mitochondrial volume fractions were foud to be 4.3 +/- 2.1% (n=52) and 1.0 +/- 1.1% (n=37), respectively.

Histochemical studies on Raillietina (Raillietina) johri (Cestoda: Davaineidae). I. Nonspecific and specific phosphatases

1979 ◽  
Vol 53 (1) ◽  
pp. 45-49 ◽  
Author(s):  
T. K. Roy
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Atpase Activity ◽  
Functional Significance ◽  
Adenosine Triphosphatase ◽  
Vas Deferens ◽  
Gland Cells ◽  
Vitelline Gland ◽  
Receptaculum Seminis ◽  
Almost All

ABSTRACTCertain phosphatases have been localized by histochemical techniques in various tissues of a pigeon cestode, Raillietina (Raillietina) johri. Acid phosphatase (AcPase), alkaline phosphatase (AlPase) and adenosine triphosphatase (ATPase) were present in almost all structures: tegument; subtegumental muscles; subtegumental cells; excretory canal; tsetes; sperm ductules; vas deferens; cirrus sac; cirrus; ovary; receptaculum seminis; vagina; vitelline gland cells; oocytes; uterus; embryonated eggs. AlPase was absent in parenchyma, spermatocytes, spermatids and spermatozoa. AlPase activity was more intense in the tegument of mature gravid proglottides. AcPase and ATPase were visualized in various stages of spermatogenesis of the parasite. ATPase activity was also observed in chromosomes. 5'-nucleotidase (AMPase) activity was restricted to embryonated eggs only. Functional significance of these phosphatases is discussed.

Parallel Intralysosomal Amyloid Fibrils, a Possible Result of Phagocytosis

1977 ◽  
Vol 14 (4) ◽  
pp. 407-419 ◽  
Author(s):  
E. Gruys ◽  
M. Castaño
Keyword(s):  
Electron Microscopy ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Enzyme Histochemistry ◽  
Amyloid Fibrils ◽  
Light And Electron Microscopy ◽  
Interstitial Cells ◽  
Mesenchymal Cells ◽  
Parallel Arrangement

Vacuoles of mesenchymal cells in the papillae of bovine kidneys with amyloidosis were studied by histochemical electron microscopy for acid phosphatase as a marker for lysosomes. The vacuoles contained parallel amyloid fibrils. The vacuoles of reticular interstitial cells were found to be lysosomes. Vacuoles of macrophage-like cells of the same papillae were positive, partially positive, or negative for the enzyme activity. A suspension of papillary material was injected subcutaneously in rats in a 21-day light and electron microscopy and enzyme histochemistry study. Amyloid was demonstrated in vacuoles of macrophages throughout this period and initially also in neutrophils. In most vacuoles amyloid fibrils were randomly arranged but in others parallel arrangement was demonstrated. Amyloid was only at the inoculation sites. Intralysosomal bovine amyloid may occur in parallel fibrillar arrangement without a definite indication for amyloid production.

scholarly journals OBSERVATIONS ON THE FINE STRUCTURE OF LUTEIN CELLS

1962 ◽  
Vol 12 (1) ◽  
pp. 101-113 ◽  
Author(s):  
Allen C. Enders
Keyword(s):  
Electron Microscopy ◽  
Endothelial Cells ◽  
Endoplasmic Reticulum ◽  
Fine Structure ◽  
Steroid Hormones ◽  
Osmium Tetroxide ◽  
Secretory Activity ◽  
Corpora Lutea ◽  
Pericapillary Space ◽  
Lutein Cell

Corpora lutea from the period of delayed implantation and from early postimplantation stages of the armadillo, mink, and rat were fixed in buffered osmium tetroxide-sucrose or potassium permanganate. After rapid dehydration, the portions of the corpora lutea were embedded in either methacrylate or epoxy resin. Examination of the lutein cells by electron microscopy revealed the presence, in the better preserved material, of an extensive development of tubular agranular endoplasmic reticulum. Although the membranes of the endoplasmic reticulum are the most striking feature of the lutein cells of both stages of the three animals examined, very numerous large mitochondria with cristae that exhibit a variety of forms tending toward villiform, and protrusions and foldings of the lutein cell margins on the pericapillary space are also characteristic of these cells. Certain minor differences in the lutein cells of the species examined are also noted. No indications of conversion of mitochondria into lipid, of accumulation of lipid in the Golgi area, or of the protrusion of lutein cells into spaces between the endothelial cells, as suggested by other authors, were noted in these preparations. Some of the difficulties inherent in the visualization of the secretory activity of cells producing steroid hormones are briefly discussed.

Ultrastructure of the Intestinal Absorptive Cells of Fed and Fasted Tadpoles

1970 ◽  
Vol 28 ◽  
pp. 86-87
Author(s):  
Robert Giaquinta ◽  
M. A. Hayat
Keyword(s):  
Electron Microscopy ◽  
Small Intestine ◽  
Fine Structure ◽  
Osmium Tetroxide ◽  
Rana Pipiens ◽  
Ultrastructural Changes ◽  
Amphibian Metamorphosis ◽  
Absorptive Cells ◽  
Food Absorption ◽  
Tissue Specimens

The ultrastructural changes that occur in the intestinal absorptive cells during amphibian metamorphosis have been reported (Bonneville, 1963). These changes accompany a change in diet (from an herbivorous to a carnivorous state) during metamorphosis. Little information is available, however, on the ultrastructural changes in the absorptive cells of amphibians in relation to the state of feeding. This report describes the differences in the fine structure of these cells in the tadpole stage of Rana pipiens during periods of food absorption and fasting.Rana pipiens at tadpole stages were fed an herbivorous diet, and after a period of 48 hr, the animal was dissected and segments of the small intestine were collected for electron microscopy. A second group of tadpoles was fasted for 7 days, and segments of the small intestine were collected. The tissue specimens were immersed in phosphate-buffered glutaraldehyde (3%) for 1 hr at 4C and postfixed with phosphate-buffered osmium tetroxide (2%) for 1 hr at 4C.

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