Just for some context, I'm isolating non parenchymal cells with Pronase+Collagenase D, which is a well known protocol in my lab. I use DAPI as a viability dye in my FACS panel but I encounter some issues as it also stains high DNA content cells or mitotic cells, representing around 20% of the cells (see picture). No solution in the protocol contains detergent that could permeabilize the cells, and the mouse used is a classic healthy C57 so mitotic cells are not expected, at least to that extent. Did anyone encountered the same issue in the past? Thank you
Thank you both for your answers, I initially used a concentration of 1.6 µg/ml concentration. By reducing by half the concentration and avoiding too long incubation time, I was able to avoid the issue !
Maxime
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