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[分离细胞涂片糖原保存的条件。II. 固定对细胞糖原含量影响的细胞荧光分析研究]

[Conditions for preserving glycogen in smears of isolated cells. II. Cytofluorimetric study of the effect of fixation on the glycogen content of cells].

作者信息

Kudriavtseva M V, Shalakhmetova T M

出版信息

Tsitologiia. 1978 Jul;20(7):772-7.

PMID:695008
Abstract

The influence of fixation on the intensity and specificity of the fluorescence of PAS-reaction (F-PAS) in the rat's liver cells was examined. 1.4 types of fixatives, routinely used in polysacchride cyto- and histochemistry, were tried: 100% metanol, 100% and 80% ethanol, acetone, 10% neutral formalin, buffered neutral formalin, mixtures of various fluids: ethanol : acetone (1 : 1), ethanol : formalin (9 : 1), ethanol : acetic acid (3 : 1), formaline : ethanol : acetic acid (1.0 : 8.5 : 0.5), fixatives of Carnoy, Bouin, Rossman and Shabadash. The cell fluorescence intensity after F-PAS reaction with Schiff's reagent auramine--SO2 and the cell autofluorescence were measured cytofluorometrically. It was shown that all the fixatives, besides Bouin's and Shabadash's fluids provide rather good preservation of the cell glycogen. Results obtained from the cytofluorometry of F-PAS reaction, autofluorescence and from the morphological studies of F-Pas stained cells, suggested that the best fixatives were 100% metanol and 100% ethanol.

摘要

研究了固定对大鼠肝细胞中PAS反应(F-PAS)荧光强度和特异性的影响。1. 尝试了多糖细胞化学和组织化学中常规使用的4种固定剂:100%甲醇、100%和80%乙醇、丙酮、10%中性福尔马林、缓冲中性福尔马林、各种液体的混合物:乙醇:丙酮(1:1)、乙醇:福尔马林(9:1)、乙醇:乙酸(3:1)、福尔马林:乙醇:乙酸(1.0:8.5:0.5)、Carnoy、Bouin、Rossman和Shabadash固定剂。用席夫试剂金胺-SO2进行F-PAS反应后的细胞荧光强度和细胞自发荧光通过细胞荧光测定法进行测量。结果表明,除了Bouin氏液和Shabadash氏液外,所有固定剂都能较好地保存细胞糖原。F-PAS反应的细胞荧光测定、自发荧光以及F-Pas染色细胞的形态学研究结果表明,最佳固定剂是100%甲醇和100%乙醇。

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