流式细胞术分选外周血细胞方法的研究

Application of sorting peripheral blood cells by flow cytometry in the detection of peripheral Mood cell sorting

目的 应用荧光激活细胞分选技术（FACS）分选T淋巴细胞（CD3^＋）和粒细胞（CD15^＋）,建立人类外周血各细胞亚群的分离方法,为基于各细胞亚群的科研和临床研究奠定实验基础.方法 收集10例健康人的外周血,利用抗体分别标定粒细胞及T淋巴细胞,应用FACS进一步分选出CD3^＋T淋巴细胞及CD15^＋粒细胞.结果 分选前外周血中CD3^＋T淋巴细胞约占白细胞总数的30.8%,CD15^＋粒细胞约占白细胞总数的48.8%;FACS分离纯化后的CD3^＋T淋巴细胞的纯度可达98%,回收率为95%;CD15^＋粒细胞的纯度可达97%,回收率约为96%.结论 应用FACS可从外周血中快速分离高纯度的CD3^＋T淋巴细胞及CD15^＋粒细胞.

Objective To apply fluo-rescencc-activated cell sorting（FACS） in sorting T lymphocyte （CD3^＋） and granulocyte （CD15^＋）, which establish the separating method of series of cells from human peripheral blood, so that the scientific research and clinical research could be carried out were specifically. Methods 10 cases of normal peripheral blood were collected and T lymphocyte （CD3^＋） and granulocyte （CD15^＋） were stained with florescence conjugated antibodies. The positive cells were sorted by FACS. Results Before sorting the peripheral blood, the proportion of the T lymphocyte（CD3^＋） and granulocyte （CD15^＋） in the leukocyte is 48.8 % and 30.8 %; after sorting by FACS, the purity of T lymphocytes （CD3^＋） is up to 98 % and the recovery is about 95 %; the purity of granulocyte （CD15^＋） is up to 97 % and the recovery is about 96 %. Conclusion The FACS could Mlow us to quickly sort T lymphocyte （CD3^＋） and granulocyte （CD15^＋） with higher recovery and higher purity from the peripheral blood.