Hello! I have been trying to increase cell density after FACS. I was sorting retina ganglion cells from adult zebrafish retina. I can get 15k cells in 40µl AMES. I put this directly to centrifuge. However, failed to increase cell density. The cell concentration across the tube after centrifuge is almost homogenized. I would be more than appreciated if someone can provide any insight on this issue. Thanks!
Fisherbrand™ 0.2mL PCR Tubes, may be spin in small size tubes, that might help you,
good luck
Dear all,
Thank you for your answers! It turns out I lost most of my cells on the centrifuge tube wall. My cells landed using the following conditions:
•30k live cells (all cell types), 500µl low adhesion tubes, 3% BSA, swing bucket centrifuge, 200 rcf, 5 min.
•100 cells/µl, viability 75% → 1000 cells/µl, viability 70%