Fixing adherent cells

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.

Remember:

• 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.