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流式细胞术测定人红细胞中的碳水化合物

Flow cytometric determination of carbohydrates in human erythrocytes.

作者信息

Duijndam W A, van Duijn P

机构信息

Department of Cytochemistry and Cytometry, Sylvius Laboratories, Leiden, The Netherlands.

出版信息

Histochemistry. 1988;88(3-6):263-5. doi: 10.1007/BF00570282.

Abstract

Carbohydrate components known from biochemical analysis to be present in peripheral normal human erythrocytes so far could not be detected cytochemically. By periodic acid oxidation followed by Schiff pararosaniline (SO2) staining, however, a specific fluorescent signal can be obtained, strong enough to allow measurement by flow cytometry. Dimethylsuberimidate fixation results in low autofluorescence and low staining of unoxidized cells. By treating erythrocyte ghosts similarly, it is found that about 20% of the signal is present in the membrane, most probably due to glycophorins. The main signal resides in the matrix of the fixed erythrocyte and may be due to traces of glycogen and to the glycosylation of proteins, especially hemoglobin.

摘要

到目前为止,通过生化分析已知存在于正常人外周红细胞中的碳水化合物成分,无法通过细胞化学方法检测到。然而,通过高碘酸氧化,然后用席夫对品红(SO2)染色,可以获得足够强的特异性荧光信号,以便通过流式细胞术进行测量。亚氨酯二甲酯固定导致未氧化细胞的自发荧光低和染色浅。通过对红细胞影进行类似处理,发现约20%的信号存在于膜中,很可能是由于血型糖蛋白。主要信号存在于固定红细胞的基质中,可能是由于微量糖原以及蛋白质(尤其是血红蛋白)的糖基化。

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