去污剂处理后可使T和B淋巴细胞在形态学上易于鉴别

niwanmao

在形态学上，T和B不易区分，但是经过去污剂4DL和4DS处理后，可使T和B淋巴细胞在形态学上易于鉴别，原理就是利用T和B两者细胞膜上的脂质含量不同。
这个内容对于一些做形态方面的，或者是流式上需要结合形态进行判断的情况，比较有帮助。不过不知道4DL和4DS这两个去污剂（Sysmex的产品）是不是单卖的？

下图显示的是去污剂处理后，T和B细胞发生的变化和差异

去污剂处理后，T和B细胞发生的变化和差异

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Non-activated T and B Lymphocytes Become Morphologically Distinguishable After Detergent Treatment

Abstract
T and B lymphocytes are difficult to distinguish morphologically even with electron microscopy, and antibodies are generally used to make the distinction. A specific reagent, consisting of nonionic and cationic detergents, is used for leukocyte differentiation using the Sysmex automated blood analyzer. This reagent increases cell membrane porosity and enables the introduction of fluorescent dye into leukocytes. In this study, we investigated the effect of this specific detergent on the morphology of T and B lymphocytes. T and B lymphocytes were obtained by density gradient centrifugation and magnetic cell sorting, with a minimum of 90% isolation efficiency. T and B lymphocytes were then treated with the specific detergent and fluorescent dye, and their distribution was analyzed based on side scatter and fluorescence intensity using general-purpose flow cytometry (FCM). Fluorescent images were observed using a confocal laser scanning microscope (CLSM), cellular inner structures using a transmission electron microscopy (TEM), and cell surfaces using a scanning electron microscope (SEM). The ratio of cholesterol
to total lipid in cell membranes of B and T lymphocytes was measured using a fluorescent assay kit. The distribution of fluorescence intensity was different between T and B lymphocyte clusters, according to the FCM analysis. CLSM observations revealed that the fluorescent dye mainly stained cytoplasmic organelles. FCM, TEM, and SEM observations revealed that B lymphocytes are more likely to lose surface antigens and intracellular organelles than T lymphocytes, which allows the visual distinction between T and B lymphocytes. The ratio of cholesterol to total lipid in T lymphocyte membranes had tendency higher than that in B lymphocyte membranes. In this study, we demonstrate that cells with differences in cell membrane cholesterol amounts, such as B and T lymphocytes could be identified using an inexpensive detergent, as an alternative to costly antibodies.

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