Cell Loss during staining
I have been having major issues with cell loss during the staining protocol. The biggest loss happens right after the washes following the permeabilization with 1X ebiosciences perm buffer. I have been staining in small 96-well plates but have also tried the 96-deep well plates but have had the same issue. Is thegre something I am fundamentally doing wrong? ALl washes prior to the 2% O/N PFA fix are done at 1600 10mins...after the PFA fix, I wpin 2000rpm 5 minutes. Any input would be appreciated!