Team:EPF-Lausanne/Protocol/FACS

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Protocol: FACS


We've used flow cytometry to check the measure the expression of LovTAP. As we had no machines in our lab, we used the FCCF's (Flow Cytometry Core Facility) machines to perform the measurements. The cells normally need to be [http://openwetware.org/wiki/Fixing_cells fixed] before being brought to the lab but, as our project has a biosafety level 1, we could bring in the cells alive.

The sample preparation required only a single washing step with PBS:

  1. Centrifuge the cells at 1500rpm for 3 minutes in a centrifuge (or at 3000rpm for 3 minutes in a microcentrifuge).
  2. Aspirate away the medium (be very careful not to disturb the pellet).
  3. Resuspend in PBS (we used ~400µl for 400'000 cells, but the required density is different for every machine).