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[检测天然血涂片白细胞中脱氢酶活性的细胞化学方法]

[Cytochemical method for detecting dehydrogenase activity in the leukocytes of native blood smears].

作者信息

Bykov E G, Chaĭtsev V G, Petrov A V, Gurevich N L

出版信息

Tsitologiia. 1987 Mar;29(3):362-5.

PMID:3296367
Abstract

A cytochemical method of detection of dehydrogenases in blood leucocytes is proposed. Native smears are dried up in the air to be incubated at 37 degrees C in gel-containing medium composed of polyvinyl alcohol, sucrose, a corresponding substrate, cofactors and inhibitors of cytochrome oxidases activity. Using corresponding media, activities of succinate, malate, glutamate, lactate-, alpha-glycerophosphate, alcohol, beta-oxybutyrate and glucoso-5-phosphate dehydrogenases were revealed. Half-reduced diformazan providing diffuse rosy staining of cells was removed after the incubation, and the incubation medium was washed out by rinsing the smears in 60% acetone solution. As a result monochromatic micropreparations may be received. Finally, smears are fixed in formalin. The above method provides a reduced loss of enzymes, preserves a good cell morphology and eliminates non-dehydrogenase effects of tetrazolium reduction into formazan.

摘要

提出了一种检测血液白细胞中脱氢酶的细胞化学方法。将天然涂片在空气中干燥,然后在由聚乙烯醇、蔗糖、相应底物、辅酶因子和细胞色素氧化酶活性抑制剂组成的含凝胶培养基中于37℃孵育。使用相应的培养基,可显示琥珀酸、苹果酸、谷氨酸、乳酸、α-甘油磷酸、乙醇、β-羟丁酸和葡萄糖-5-磷酸脱氢酶的活性。孵育后去除提供细胞弥漫性玫瑰色染色的半还原双甲臜,通过在60%丙酮溶液中冲洗涂片洗去孵育培养基。结果可得到单色显微制片。最后,涂片用福尔马林固定。上述方法减少了酶的损失,保持了良好的细胞形态,并消除了四氮唑还原为甲臜的非脱氢酶效应。

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