Zvetkova E B, Zvetkov I B
Acta Histochem. 1976;57(1):1-13. doi: 10.1016/S0065-1281(76)80002-5.
A cytochemical method is suggested for the simultaneous and differential staining of cellular nucleoproteids [ribonucleoproteids (RNP) and desoxyribonucleoproteids (DNP)], as well as for the simultaneous contrast staining of some basic (arginine- and lysin-containing) proteins. The staining technique is based on DNA-denaturation procedures and the application of mixtures of basic dye--methylene blue and acid dyes--eosin or fast green at low concentrations. The combination of methylene blue with eosin is used for the staining of ribonucleoproteids (RNP) whereas methylene blue-fast green for the simultaneous detection of ribonucleoproteids and desoxyribonucleoproteids (RNP and DNP), as well as for the differential staining of nuclear DNP (after cold hydrolysis with 5 N HCl). The acid dyes eosin and fast green stain in pink resp. in green some cathionic proteins in the lysosomal (specific) granules of the neutrophilic and eosinophilic leucocytes in the cytoplasm of erythrocytes, and after cold hydrolysis in the cytoplasm of lymphocytes. A fluorescent variant of the method with sulfaflavin is also suggested for the fluorochromation of cytoplasmic cathionic granules in the luecocytes. Acid mucopolysaccharide components in the granules of basophilic leucocytes, tissue mastocytes and thrombocyres are stained intensively pink-violet (gamma-metachromatic). The possibilities for the application of the method in the quantitative analysis of blood and exfoliated cells, as well as for purpose of haematology, immunology and exfoliative cytology are discussed.
本文提出了一种细胞化学方法,用于对细胞核蛋白(核糖核蛋白(RNP)和脱氧核糖核蛋白(DNP))进行同时和鉴别染色,以及对一些碱性(含精氨酸和赖氨酸)蛋白质进行同时对比染色。染色技术基于DNA变性程序以及低浓度碱性染料(亚甲蓝)和酸性染料(伊红或固绿)混合物的应用。亚甲蓝与伊红的组合用于核糖核蛋白(RNP)的染色,而亚甲蓝 - 固绿用于同时检测核糖核蛋白和脱氧核糖核蛋白(RNP和DNP),以及用于核DNP的鉴别染色(用5N HCl冷水解后)。酸性染料伊红和固绿分别将红细胞细胞质中嗜中性和嗜酸性白细胞溶酶体(特异性)颗粒中的一些阳离子蛋白染成粉红色和绿色,在淋巴细胞细胞质中冷水解后也是如此。还提出了一种使用磺胺黄素的该方法的荧光变体,用于白细胞细胞质中阳离子颗粒的荧光染色。嗜碱性白细胞、组织肥大细胞和血小板颗粒中的酸性粘多糖成分被强烈染成粉紫色(γ-异染性)。讨论了该方法在血液和脱落细胞定量分析以及血液学、免疫学和脱落细胞学中的应用可能性。