实验概述】
本实验成功采用FACSCelesta的10色方案进行人外周血白细胞中T细胞亚群的的免疫分型，实现了一管即清楚区分Th/Ts/Treg/TEM/TCM/NaïveT/Mature T等十几个T细胞亚群，对于T细胞亚群的成熟、分化及活化等的研究起到重要作用！
【实验方法】
①/ 仪器配置
| Flow cytometer | Laser |
| FACSCelesta | 488nm Blue Laser 640nm Red Laser 405nm Red Laser |
②/ 多色配色方案及流式试剂
● 配色方案
| PE | APC-Cy7 | Alexa Fluor-700 | BV421 | BV510 | BV605 | PE-CF594 | PerCP-Cy5.5 | BB515 | BV786 |
| CD127 | CD3 | CD45RA | CD25 | CD8 | CD4 | CCR7 | CD45 | CD28 | CD27 |
● 流式试剂
| Products Name | Catalog No. | Clone | Size |
| BD Pharmingen™ PE Mouse Anti-Human CD127 | 557938 | HIL-7R-M21 | 0.1 mg |
| BD Pharmingen™ APC-Cy™7 Mouse Anti-Human CD3 | 557832 | SK7 | 100T |
| BD Pharmingen™ Alexa Fluor® 700 Mouse Anti-Human CD45RA | 560673 | HI100 | 50T |
| BD Horizon™ BV421 Mouse Anti-Human CD25 | 562442 | M-A251 | 100T |
| BD Horizon™ BV510 Mouse Anti-Human CD8 | 563919 | SK1 | 100T |
| BD Horizon™ BV605 Mouse Anti-Human CD4 | 562658 | RPA-T4 | 100T |
| BD Horizon™ PE-CF594 Mouse Anti-Human CD197 (CCR7) | 562381 | 150503 | 50T |
| BD Pharmingen™ PerCP-Cy™5.5 Mouse Anti-Human CD45 | 564105 | HI30 | 100T |
| BD Horizon™ BB515 Mouse Anti-Human CD28 | 564492 | CD28.2 | 100T |
| BD Horizon™ BV786 Mouse Anti-Human CD27 | 563327 | L128 | 100T |
| BD Pharm Lyse™ Lysing Buffer (10x ) | 555899 | 100ml |
【实验结果】
如图所见，所有群体阴阳性区分明显，群体集中，即使是弱表达群体Treg也区分非常清楚，实验中使用了Diva软件的自动补偿功能，补偿准确、操作极其简单，为多色实验提供方便、快捷且精确的补偿操作。
* 本资料仅限科研用途使用
