Hello!
I am doing a bit of sorting recently. It used to be okay - approximately 90% purity. Unfortunately, yesterday it was a kind of random. The total number of cells in collection tube was fine, but they were totally not sorted. I observed similar results for Th cells and monocytes. For example, in monocyte sorting (three subsets), in each collection tube we had mostly lymphocytes and up to 5% monocytes.
We used the 100um nozzle, 20psi pressure, the gap was 14, 1st drop 314. Drop delay was set automatically on those settings. The stream was stable. Event rate approximately 1200/s.
Do you have any suggestions? What am I doing wrong?
Thank you!
I know this is an old question but I had the same problem today and found a solution. For whatever reason, it seems that the software on my machine calculated the drop delay incorrectly. This resulted in the machine being unable to correctly sort different cell populations (pretty much everything ended up in the sort tubes). Re-running the auto drop-delay using the same tube of Accudrop beads corrected the issue and I was able to sort samples as normal.
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