Guimaraes J E, Berney J J, Francis G E, Hoffbrand A V
Exp Hematol. 1984 Aug;12(7):535-8.
The identification of mixed granulocytic-erythrocytic colonies in culture poses special problems. First, methods based on the detection of peroxidase-like activity of red-cell hemoglobin using benzidine can give false-positive results with eosinophilic cells in colonies. Secondly, the use of the color of hemoglobin itself to identify mixed colonies prevents the detection of those colonies where the erythrocytic component is small or contains relatively little hemoglobin. We present here a new method using O-Dianisidine (O-D) to detect red-cell peroxidase activity. Subsequent staining with Luxol fast blue showed that eosinophils did not give a positive reaction with O-D. The technique also has the advantage that the whole culture gel is processed and that there is no need to pick off individual clones for identification. Using the combined stain O-D and Luxol fast blue most lineages within clones can be discriminated.
在培养物中鉴定混合性粒细胞-红细胞集落存在特殊问题。首先，基于使用联苯胺检测红细胞血红蛋白类过氧化物酶活性的方法，可能会在集落中的嗜酸性细胞上产生假阳性结果。其次，利用血红蛋白本身的颜色来鉴定混合集落，会妨碍对那些红细胞成分少或血红蛋白含量相对较低的集落的检测。我们在此介绍一种使用邻联茴香胺（O-D）检测红细胞过氧化物酶活性的新方法。随后用卢戈氏坚牢蓝染色表明，嗜酸性粒细胞与O-D不产生阳性反应。该技术还有一个优点，即对整个培养凝胶进行处理，无需挑选单个克隆进行鉴定。使用O-D和卢戈氏坚牢蓝的联合染色，可以区分克隆内的大多数谱系。
