Shreffler:CFSE Labeling

Materials

CFDA/SE Invitrogen C1157 as 5 mM stock in DMSO

Procedure

Suspend PBMCs at 10x10⁶ cells/mL in PBS alone.

Cells should be in PBS for CFSE labeling.

If CFSE negative control is needed, remove cells now.

Make '2X' concentration (10 μM) in PBS

• Add 4mL PBS + 8 μL 5 mM CFSE (for example)

Combine 1:1 PBMC cells + CFSE (i.e. 300uL PBMCs + 300uL CFSE) in 15 ml tube.

Place in H₂O bath x 10 min.

Wash in 10 mL complete medium (10 min at 300G). Aspirate.

Resuspend in medium at desired density