细胞计数板在荧光显微镜观察B细胞吞噬现象的应用探讨

The Explore of Phagocytosis of B Lymphocytes using Cell Counting Chamber under Immunofluorescence Microscopy

目的:基于细胞计数板建立一种简单、快速使用免疫荧光显微镜观察B淋巴细胞吞噬卡介苗(BCG)现象的新方法,对即将进行流式细胞检测的样品进行质控,提高流式细胞术检测吞噬率的稳定性,同时为流式细胞仪检测吞噬率提供镜下依据。方法:B细胞与FITC标记的BCG共培养24 h后,PE anti-human CD19抗体直接标记细胞膜,应用细胞计数板在荧光显微镜下观察B细胞吞噬现象,流式细胞仪检测吞噬率。结果:应用细胞计数板在荧光镜下可观察到B细胞与BCG的荧光标记及B细胞与BCG共标记现象,证实B细胞可吞噬BCG,流式细胞仪检测结果显示吞噬率为13.9%。结论:应用细胞计数板在荧光镜下可观察B细胞吞噬现象,且操作简便快速,能对流式细胞检测的样品进行质控,并提供镜下依据。

Objective： To establish a simple and quick method to explore phagocytosis of B lymphocytes under immunofluores- cence microscopy based on cell counting chamber. By monitoring the sample quality before flow cytometry（FCM）, the method could im- prove the stability of phagocytic rate detected by FCM. It could also offer image proof under fluorescence microscopy at the same time. Methods： After Raji B lymphocytes and FITC-stained BCG were incubated together for 24 hrs, B lymphocytes were labeled by PE an- ti-human CD19 antibody. Then we applied cell counting chamber to observe phagocytosis of B lymphocytes and flow cytometry to test phagocytic rate. Results： B lymphocytes and BCG could be single labeled respectively and double labeled,which was observed under flu- orescence microscope based on cell counting chamber. The phagocytic rate tested by flow cytometry was 13.9 %. Conclusions： Observ- ing phagocytosis of B lymphocytes using cell counting chamber under fluorescence microscopy might be a simple and quick method. It can be applied in monitoring the sample quality before flow cytometry, it could also offer proof under fluorescence microscopy.