Oil Red O staining

1. Rinse cells two times with PBS
2. Fix cells in 1mL/6-well 10% formalin/PBS for one hour (no more than 2 hours) at RT.
3. Rinse plates two times with ddH₂O
4. Stain with 1mL/6-well Oil Red O (stain prep below) for 2 hours at RT.
5. Aspirate stain and wash cells gently with water a many times (around 15-20, until background stain comes off).
6. Stained fat droplets will be dark brown/red in color.
7. Leave some ddH2O on the plates and image.
8. Either keep moist with PBS or air dry.

Preparation of Oil Red O:

1. 1.05g Oil Red O dissolved in 300mL isopropanol and leave overnight at RT w/o stirring
2. Filter with 0.45um membrane, add 250mL ddH₂O and leave solution overnight at 4°C without stirring
3. Filter 1x with 0.45um filter
4. Filter 1x with 0.22um filter
5. store at RT