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[single_cell_nerd_88]

​Hi,

 

​I had a question for the Flow experts here. Has anyone tried staining with multiple dyes - 7-AAD, Annexin-V (PE) - part of Annexin V Apoptosis Detection Kit from BD Pharmingen. These are to be coupled with Dyecycle Violet (BV421) and Cell Trace CFSE (FITC). I have the following problems -

 

1) 7-AAD seems to be bleeding into the PE channel. I've tried compensating and I have been unable to do so

2) Also Annexin V (PE) is bleeding into the Dyecycle channel. 

​3) Cell Trace is bleeding into Annexin V and 7-AAD channels. I've tried knocking them down.

 

I have already tried compensation. Automatic and manual. I am running my samples on the FACS-Canto-II. 

 

​Has anybody else tried this combination or has experienced similar issues?

 

Thanks