high self fluorescence when doing FACS - (Sep/16/2007 )
I recently ran a flow cytometry of my rat alveolar macrophages. Even the cells only got a high background and the reading is about the same as the cells with my fluorecent labelled LPS. Does any one help me figure out the problem??
Does anyone know how to make the clumped cells as scattered as they can? This has been said might give high auto fluorescent.
If these are fixed cells, you may want to lower your PFA concentration to reduce autofluorescence. To reduce clumping, vortex vigorously after fixation and filter your cells before you run them.
These are not fixed cells. I just treat them and then run them through Flow... Thanks for the suggestion about vortexing. Certainly will do it to see if anything improves.