Yeast cell fixation with formaldehyde - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki

1. Spin down cells in eppendorf and remove supernatant
2. Add 100 microliters of 4% formaldehyde and vortex
3. incubate at RT from 15min to 1h
4. Spin cells and remove supernatant
5. Wash once in PBS (0.5-1 mL).
6. resuspend in desired volume of PBS.
7. Store cells in refrigerator for up to one month or in the freezer for even longer. The cells are now ready to be used in cytometry.

4% formaldehyde

1. Dissolve 2 g of paraformaldehyde in 40 mL of warm PBS. NOTE: formaldehyde is bad for you, so cover the top of the beaker while you mix this up.
2. Bring the volume up to 50 mL with PBS
3. Store at room temperature

1X PBS

1. To 800 ml of distilled water add:

• 8 g of NaCl,
• 0.2 g of KCl;
• 1.44 g of Na2HPO4;
• 0.24 g of KH2PO4.

2. Adjust the pH to 7.4 with HCl. Add distilled water to a total volume of 1 liter.