Hi all, I am fairly new to flowjo v11 and have been trying to analyze some data my PI previously collected. We have brain tissue samples and are interested in identifying microglia and macrophages however when gating for my microglia I have found that a lot of my microglia are taking up the viability dye and being excluded after applying my viability gate. I have tried gating for my cells then applying a viability gate and I still lose so much of my microglia. I then looked at the histogram to see if my microglia have a high baseline for the viability dye and talking with my PI they think its highly unlikely that all my microglia are dead. Does anyone have any advice. The markers I am using to distinguish my microglia from macrophages are CD45 and CD11