I want to test whether there is difference in the in vivo cell migration in WT and PST-/- myeloid cells in mice model. For this, I have been using CellTracker™ Deep Red (catalog no. C34565) and Vybrant™ CFDA SE Cell Tracer (catalog no. V12883) to label WT and PST-/- bone marrow cells respectively. Cells were labeled separately then mixed to do the competitive study. before injecting them into mice I went to confirm the labeling and separation on these two cells. To my surprise we found that cells got dual staining. We have repeated this several times. We did modifications like incubating cells separately from 1hr to over night to make the staining stable, changing concentration, increasing number of washes etc.

But we got same outcome, instead of getting two different cell clusters labeled either with deep red or CFDA- SE we got dual stained cells.

All experiments were done as per the company protocol. Used only PBS and RPMI+ 10%FBS.