GFP and PE
Posted 01 March 2011 - 07:40 AM
I work on GFP positive cells out from a transgenic mouse, expressing GFP in a certain celltype.
If I do flow-cytometry analysis with these cells I canīt see GFP positive cells in combination with PE.
APC seems to be the better combination - but then I just have one colour left (if working on FACSCAlibur).
Does anybody has experiences staining GFP+ cells with PE or PerCP or other colours (that can be read with the 488nm or 635 nm laser)
Thank you very much !!!
Posted 01 March 2011 - 11:39 PM
If you use ToPro3 for DNA (APC channel), works very well as well.
Posted 02 March 2011 - 01:46 AM
I think because GFP glares quite bright into the FL2 channel.
With control stains (FITC & PE) I can compensate both channels quite well - but if measuring GFP cells I get an FL2 population from the GFP cells (with the same adjustment as the control) If I compensate the "false-positive" FL2 population, PE positive cells and as well the GFP population are not detectable anymore. (they fall into the negative)
Just I got the information that GFP and PE can not be combined - but some people can do it...?
Posted 03 March 2011 - 12:06 AM