I have started running samples for a research project where the investigators label mouse lymphs with either CFSE or transfect with GFP and then transplant into another mouse. After 1 to 7 days they remove tissues from the recipient mouse (blood, spleen, thymus, lymph node) and try to detect the donor cells. These events can be very rare (<1%). They are also labelling two other surface markers, so it is three color staining. Any tips on doing this right would be greatly appreciated. I am most concerned about the number of events to collect and what to use for a positive control for CFSE/GFP for compensation. Thanks in advance. David McFarland Howard Hughes Medical Institute Flow Cytometry Facility Vanderbilt University Medical Center
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