Journal of Histochemistry & Cytochemistry

FISHMAN, JOEL S.; HAYASHI, MASANDO

doi: 10.1177/10.5.515pmid: N/A

The enzymorphology technique with chloral hydrate fixative as applied to rat brain gives good preservation of the cell morphology and enzyme activity. The findings are in harmony with those obtained by others using separate techniques. Thus, β-glucuronidase shows an intense activity in the Purkinje cells of the cerebellum; α-naphthyl esterase and indoxyl esterase show strong activity in the neurons, astrocytes and their processes and alkaline phosphatase enriched the walls of the capillaries.

FREIMAN, DAVID G.; GOLDMAN, HARVEY; KAPLAN, NANCY

doi: 10.1177/10.5.520pmid: N/A

Pretreatment of cryostat sections of dog kidney with sodium salts of various anions in acetate buffer at pH 4.0 for periods of time ranging up to 6 hours produces differential inhibition of non-specific alkaline phosphatase activity as determined by the Gomori method. The degree of anionic effect is dependent on the nature of the anion and modified by the cation present. The effect also varies directly with the anionic concentration, duration of exposure and temperature of the pretreatment buffer and inversely with the pH. On the basis of the differential suppression of enzymatic activity by the various anions, and the conditions under which these effects were evident, distinctive inhibition profiles were established for the several phosphomonoesterases tested, including the renal and intestinal alkaline phosphatases and the smooth muscle 5'-nucleotidases of several species. The similarity of the results obtained in various biochemical studies on other enzymes reported in the literature and those obtained histochemically in these experiments suggests a common mechanism of action; this is presumed to be a differential effect of anions on the dissociation of apoenzyme and coenzyme. The method described provides a simple and effective means of differentiating, under histochemical conditions, closely related and even apparently identical enzymes.

SPICER, S. S.

doi: 10.1177/10.5.528pmid: N/A

Prussian blue-reactive particles, designated cytosiderin, have been demonstrated in various epithelia of rats, hamsters and guinea pigs retired from the breeding colony. Weanling animals lack such deposits. The distribution of cytosiderin in the various histologic sites differs for each species. Morphologically similar particles have been visualized by means of staining with oxidation-aldehyde fuchsin techniques. The latter cytoplasmic particles, present in the rodents examined as well as in human tissues, were encountered either in the same organs as those with cytosiderin or in organs lacking Prussian blue-reactive particles.

ZERLOTTI, EUGENIO; ENGEL, MILTON B.

doi: 10.1177/10.5.537pmid: N/A

2,4-Dinitrofluorobenzene was used without any coupling procedure as a histochemical reagent to localize some reactive groups of proteins in connective tissues and epithelial structures. Tissues were fixed by freezing-drying to avoid uncontrolled alterations of proteins and the reactions were modified in some sections by postfixation or pre-treatment with a variety of reagents, including alcohol, formalin, formaldehyde vapors, acetic anhydride, nitrous acid and N-ethylmaleimide. The staining and resolution were greatly enhanced by viewing the sections with monochromatic light at 410 mµ. When used in this way, 2,4-dinitrofluorobenzene is specific for: (1) α-amino groups of terminal amino acids; (2)ε-amino groups of lysine and hydroxylysine; (3) sulfhydryl groups of cysteine. However, the reaction seems to be dominated by the ε-amino groups of lysine in most of the tissues. In the native state not all functional groups of tissue colloids react presumably because the groups are masked by the interractions involved in forming tertiary structures or covalent bonds. This is specially notable in calcified tissues. Pyknotic nuclei, pre-dentin, carious dentin and newly formed bone are more reactive suggesting a less highly organized state of their structural proteins or some other modification in the lysine and hydroxylysine containing proteins. 2,4-Dinitrofluorobenzene is valuable as a specific cytochemical reagent and, in some instances, as an indicator of the state of organization of cellular and extracellular colloids.

KASTEN, FREDERICK H.; KIEFER, G.; SANDRITTER, W.

doi: 10.1177/10.5.547pmid: N/A

Feulgen-stained cells were placed on microscope stages and exposed to monochromatic (570 mµ, 500 mµ) and polychromatic visible light in two different cytophotometers for as long as 113 hours. A bleaching effect was measured in 8 of 11 differemst experiments by scanning and multiple plug cytophotometry. Dye extinctions were reduced by 27 to 81%, varying with the particular slide preparation and conditions of irradiation. The absorption curve is also altered in shape as a result of the light exposure although the absorption maximum remains at 570 mµ. The bleaching effect was found with batches of pararosaniline or basic fuchsin from three different companies. The effect was observed in freshly-stained sections as well as in an old section. Acetic-alcohol fixation may offer some protection against bleaching compared with formalin fixation. The rate of bleaching is rapid at first and then slows down after about three days. It is thought that one or more leuco compounds are produced in this process. Since as little as 2% bleaching occurs after one hour of irradiation, the phenomenon is not considered as a source of major error in routine cytophotometry. Fronn the practical side, it is suggested that light be blocked from the slide during non-measuring periods and that irradiation time be reduced. The gallocyanin-chromalum-DNA complex is not bleached from cell nuclei after prolonged irradiation although the absorption curve is shifted to lower wavelengths. Protection against bleaching is probably afforded by the screening action of the inorganic salt.

KUGLER, J. H.; WILKINSON, W. J. C.

doi: 10.1177/10.5.556pmid: N/A

The in vitro formation of glycogen from d-glucose-C14 has been studied by histochemical and autoradiographic techniques. The incorporation of the radioactive glucose into acid soluble glycogen lends support to the suggestion that this is the only glycogen fraction that is demonstrated histochemically. These experiments also confirm the validity of the histochemical tests for glycogen.

COOK, S. F.; EZRA-COHN, H. E.

doi: 10.1177/10.5.560pmid: N/A

MEEK, EDWARD S.

doi: 10.1177/10.5.564pmid: N/A

The quantitative staining response to the naphthol yellow S technique described by Deitch (1955) has been determined on normal mammalian red blood cells using scanning microdensitometry. It is concluded that these cells can be used as a standard of reference for this type of protein when making comparisons between various types of cell populations.The possibility of mutual interference following the application of both the Feulgen and naphthol yellow S techniques to the same preparation has been investigated using mouse ascites tumour cells and leucocytes. In this material it was found that the Feulgen method led to the loss of binding sites for naphthol yellow S, but that the application of the latter did not result in the extraction of deoxyribonucleic acid (DNA). By taking naphthol yellow S absorption readings before applying the Feulgen technique, this difficulty can be avoided, and both the protein and DNA complements of individual cells can be measured without error through loss of either component.

POMERANZ, Y.

doi: 10.1177/10.5.568pmid: N/A

Young and old cultures of 12 yeasts have been stained by the Barrnett-Seligman histochemical procedure for localization of sulfhydryl groups. Thiol groups have been found distributed in the cytoplasm, but not in the cell wall. The intense staining of sulfhydryl groups by the specific stain was clearly showns in the slender thread of protoplasm between bud and mother cell.The intensity of staining in the proliferating cell was higher than in the resting cell; variations in thiol content between various yeast species were recorded. No decrease in thiol content of yeast spores, as compared with vegetative cells, has been found in Schizosaccharomyces octosporus, whereas the staining of yeast spores in Saccharomyces and Hansenula was found to be negligible. Implications of these findings are discussed.

TAKEUCHI, TADAO; TADOKORO, NOZOMU; IDE, HIDEMARO

doi: 10.1177/10.5.572pmid: N/A

Polysaccharides of glycogen class were histochemically synthesized from glucose-1-phosphate at pH 5.7 in various blood and bone marrow cells except the crythrocytic series. The intense activity of phosphorylase was indicated blue with iodine which stained a long straight chain of 1,4-linkages, while the branching enzyme (amylo-1,4 → 1,6-transglucosidase) activity was indicated red violet by staining shorter chains of a branched polysaccharide. These reactions appeared most frequently in neutrophilic, pseudoeosinophilic or amphophil series, particularly in the mature cells of the series. The histochemical synthesis of glycogen from glucose-1-phosphate in neutrophil leucocytes in the peripheral blood varies under pathologic conditions.