To fix the cells grown on a coverslip
- Work in a fume hood
- Gently rinse cells 2X in warm PBS (remove medium, add PBS, repeat)
- Leave cells in this PBS—until you add the fixative.
- Remove final PBS rinse, add about 1 mL of fixative to the cells
- Fix cells in hood for 10 mins.
- Rinse cells by dunking each coverslip in 3 beakers of PBS-Tween-Azide, 10 X per beaker.
- Place coverslip back in dish with fresh PBS-Tween-Azide
- If you are not staining immediately, store cells in PBS-Tween-Azide in fridge.
- Work in the fume hood
- Wear gloves!
- Keep track of which side of the coverslip the cells are on
- Discard used fixative in the appropriate hazardous waste container in the hood.