- Put coverslip with adherent cells in the sample chamber and wash with PBS. This involves gently spraying the PBS on the coverslip with a pippette, and gently rocking the sample chamber back and forth. This should only require about 0.5 mL of PBS.
- Aspirate PBS and fill chamber with 1 mL of 4% formaldehyde. Let sit for 15 minutes at room temperature (in cell prep box, covered with a chem wipe).
- There are variances in both time and temperature (some incubate rather than using room temperature), so this is something that could be varied if desired.
- Aspirate formaldehyde and wash twice with PBS. Aspirate PBS.
- For measurements, fill with HBSS at 37C as is typical with live cell measurements.
Berger research group at the University of Rochester