第三代流式细胞分选仪及96孔板分选单个细胞的方法及参数优化

被引：9

作者：

闵智慧 ^{[1
,2
,3
]}

程韵枫 ^{[1
,4
,2
,5
,3
]}

机构：

[1] 复旦大学附属中山医院临床研究院实验研究中心

[2] 复旦大学附属中山医院青浦分院实验研究中心

[3] 上海市器官移植重点实验室

[4] 复旦大学附属中山医院血液科

[5] 复旦大学附属中山医院青浦分院血液科

来源：

中国临床医学 | 2016年 / 23卷 / 06期

关键词：

单细胞分选; 流式细胞分选仪; 96孔板; 喷嘴; 液滴延迟;

D O I：

暂无

中图分类号：

R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

目的:用第三代流式细胞分选仪比较不同直径喷嘴分选的单细胞的得率及细胞活性,进一步完善单细胞微孔板分选的参数设置及条件优化。方法:用FACSAriaⅢ流式细胞分选仪及96孔板对Raji细胞、A549、DT 40、RAW 264.7细胞系进行单细胞分选,分别采用70μm、100μm和130μm喷嘴分选,比较分选后单细胞得率及克隆形成率。结果:在单细胞分选模式下,用3种不同直径喷嘴及96孔板分选后的单细胞得率为86.55%93.44%,即96孔板有单细胞的孔数为84～92孔;分选后的单个细胞培养7d后,经70μm、100μm和130μm喷嘴分选的单细胞克隆孔数分别为31～52孔、49～70孔、58～78孔。结论:进行单细胞微孔板分选时,在精确调节及优化实验参数的前提下,为了保证单细胞的活性,应优先选择100μm或130μm喷嘴。

引用

页码：846 / 850

页数：5

共 9 条

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