I am setting up an antigen proliferation assay using Monocytes Derived Dendritic pulsed with KLH in co-culture with lymphocytes isolated by elutriation technique. I perform a CD4 negative selection using MACS beads, then I stain with CSFE or CPD and co-culture them with DCs.

The problem is that after CFSE or CPD staining I lose 50%-60% sometimes even 80% of the CD4 T-cell. I am staining 20 M CD 4 T-cell with 5 uM CFSE or CPD, does anybody have the same problem?

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