[Cytometry] CD45-PE staining disappeared

Gareth Howell G.J.Howell at leeds.ac.uk
Fri Jul 3 04:58:16 EDT 2009


Hi
I had an alarming occurrence the other day which I can't get my head around.
I was sorting cells on the FACSAria from dental pulp stained with CD34-FITC and CD45-PE (both antibodies from BD used as instructed in the protocol). During the sort all the cells looked fine, but upon analysis my CD45-PE positive population appeared completely negative  X-(
My CD34 population was fine. I was using a low sort set-up (20psi, 100um nozzle).
Has anyone seen this happen before? I think my buffers are fine (PBS with 5mM EDTA, 25mM HEPES, pH was 6 but the 'customer' is going to ensure it's at 7 next time!).
Any ideas?
Thanks
Gareth




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