| 吴梦丽,周林甫,石晓丹,刘学武,史 明,冯国栋,赵 钢.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[J].,2017,17(11):2037-2040 |
| 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 |
| 投稿时间：2016-07-16 修订日期：2016-08-12 |
| DOI：10.13241/j.cnki.pmb.2017.11.009 |
| 中文关键词: 细胞计数板 荧光显微镜 B细胞 卡介苗 |
| 英文关键词: Cell counting chamber Immunofluorescence microscopy B lymphocyte BCG |
| 基金项目:国家自然科学基金项目（81371334） |
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| 中文摘要: |
| 摘要 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。 |
| 英文摘要: |
| ABSTRACT 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 %. Conclusion: 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. |
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