请教CFSE和GFP如何在流式上区分开？

wzr19990923

倪老师：您好！
           请教一个问题：“Decrease of Foxp3+ Treg Cell Number and Acquisition of Effector Cell Phenotype during Lethal Infection”一文中进行抑制功能实验的protocol如下：
Suppression Assay
CD4+eGFP- T cells were labeled with 1.25 mM CFSE (Invitrogen) in HBSS (Mediatech Inc.) for 15 min at 37℃. Cells were washed twice in media containing 10% FBS. 5×104 CFSE-labeled CD4+eGFP- T cells were cultured per well of a 96-well U-bottom plate with 0.5 mg/ml of a-CD3 mAb (BD Bioscience) and in presence of 105 irradiated T-depleted spleen cell feeders and decreasing amounts of CD4+eGFP+ cells (at indicated ratio) during 3 days.
（实验鼠是Foxp3eGFPreporter mice)
我的问题是：GFP好像和CFSE是同一个荧光通道的，它们是怎么区分开的呢？是根据荧光强度吗？

niwanmao

是啊，我也跟你一样的想法。
不过看他们的图上，没提到egfp+细胞的排除，我估计在培养过程中，这类EGFP＋的细胞应该是逐渐消失掉的，可能是跟饲养层细胞一样的性质。

wzr19990923

谢谢您。可能如此吧。