Tas J
Histological Laboratory, University of Amsterdam.
Acta Histochem Suppl. 1977;Suppl 18:95-100.
A qualitative microspectrophotometric detection method for heparin in situ has been developed, using data obtained previously with a model system of polyacrylamide films containing pure glycosaminoglycans (Tas & Roozemond 1973, and Tas 1975). This technique, based on the unique metachromatic properties of heparin with Toluidine Blue O in glycerol, has been worked out with rat peritoneal and mesenteric mast cells. After equilibration of the stained, air-dried cells in glycerol (for some days), the specific peak of the heparin-Toluidine blue O complex (as found in the model experiments at about 515 nm) could be recorded. It was found that the method can be used to detect unequivocally the presence of heparin in cells, even if they also contain up to 75 mole percent of other, lower sulphated-glycosaminoglycan. Otherwise the method might yield some information about the degree of sulphation of the heparin concerned. With this technique the presence of heparin has been proved in young and adult rat mast cells and for the first time now directly in normal human mast cells and normal human basophilic granulocytes.
利用先前在含有纯糖胺聚糖的聚丙烯酰胺薄膜模型系统中获得的数据（Tas和Roozemond，1973年；Tas，1975年），已开发出一种定性的原位微分光光度法检测肝素的方法。该技术基于肝素与甘油中的甲苯胺蓝O的独特异染特性，已在大鼠腹膜和肠系膜肥大细胞上进行了研究。在甘油中对染色并风干的细胞进行平衡（数天）后，可记录肝素-甲苯胺蓝O复合物的特定峰（如在模型实验中约515nm处发现的）。结果发现，该方法可明确检测细胞中肝素的存在，即使细胞中还含有高达75摩尔百分比的其他低硫酸化糖胺聚糖。否则，该方法可能会提供有关相关肝素硫酸化程度的一些信息。通过这种技术，已在幼年和成年大鼠肥大细胞中证明了肝素的存在，并且现在首次直接在正常人肥大细胞和正常人嗜碱性粒细胞中证明了肝素的存在。
