液氮保存细胞表面抗原的流式细胞仪分析

Flow Cytometric Analysis of Surface Antigens on Liquid Nitrogen-stored Cells

在免疫学研究中,用间接免疫荧光技术检测细胞表面抗原是鉴别细胞种类及细胞亚群分析中常用的方法。近年来,用流式细胞仪替代荧光显微镜以检测细胞表面抗原日益增多。但在临床样品检测中常因仪器或其它原因,不能立即检测,而需把细胞(如外周血单一核细胞)先贮存于液氮中.但液氮冻存细胞常使部分细胞死亡.由于荧光素标记物能穿过死亡细胞壁进入细胞,造成假阳性,使测得值不够准确。因此,如何排除死亡细胞的干扰,成为用流式细胞仪检测液氮冻存细胞表面抗原的一个重要问题。

A high non-specific fluorescence background imposes serious interference on a flow cytoneter in surface antigen assay for those cells stored in liquid nitrogen. The non-specific fluorescence comes from dead and dying cells, the per.neability of whose membrane has been increased so that the fluorescence reagent can enter into the cells. To solve this problem, liquid nitrogen-stored PBMNC and Raji cells were screened on different gatings for surface antigen stained with fluorescence. It was observed that the high fluorescence background as well as debris can be gated out by means of gating on FALS, ∠90°∠ and PRFL. As a result, the percentage of surface antigen positive cells determined was close to that assayed in fresh samples.