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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 rapid method of using immunofluorescence microscopy to observe the phenomenon of BCG engulfment in B lymphocytes based on the cell counting plate, to control the quality of samples to be detected by flow cytometry and to improve the detection of phagocytosis by flow cytometry Rate stability, while phagocytosis rate for flow cytometry to provide a mirror basis. Methods: After cultured with B cells and FITC labeled BCG for 24 h, PE anti-human CD19 antibody directly labeled the cell membrane. The phagocytosis of B cells was observed under a fluorescence microscope with a cell counting plate. The phagocytosis rate was detected by flow cytometry. Results: The fluorescence labeling of B cells and BCG and the co-labeling of B cells with BCG were observed under the fluorescence microscope. The results showed that B cells could engulf BCG. The results of flow cytometry showed that the phagocytosis rate was 13.9%. CONCLUSION: The phagocytosis of B cells can be observed under the fluoroscope with a cell counting plate, and the operation is quick and easy. The quality control of the flow cytometry samples can be carried out and the basis of the microscope can be provided.