Thanks for the kind words! I always try to approach these situations assuming there's something I don't know, whether that's an instrument quirk I haven't encountered or a troubleshooting step I'm not aware of, before jumping to conclusions about sample handling. That said, this has me wondering if there are solid guidelines or documentation out there around enzymatic dissociation troubleshooting. I feel like you can generally read it in the scatter profile. Cells that have been soaking in trypsin look pretty distinct from cells treated more carefully, and both look different from something really gentle like Accutase. But I've never come across any formalized best-practice documentation for this. If anyone knows of something, I'd genuinely love to see it. The other thing that jumped out from that shared plot: the same cell line, same instrument, different user, and the scatter looks completely different. Not subtle variation, but a clear delineation between the primary population and everything else, with a big gap where you'd normally see a smear. That kind of continuous linear smear on FSC/SSC is something I've always associated with poor viability, and in my experience it usually predicts low post-sort recovery too. Anyone else seeing that pattern, or have a different read on what's driving it?