Wright’s Staining Protocol for Millicell®

Materials

  • Millicell®-CM cell culture plate inserts — Millipore cat. nos. PICM01250, PICM03050
  • 3% glutaraldehyde in Phosphate Buffered Saline (PBS)
  • Milli-Q® water
  • 100% methanol
  • Wright’s stain

Method

  1. Remove the Millicell®-CM insert from the plate and wash gently with PBS to remove growth media.
  2. Fix the cells for 15 minutes with 3% glutaraldehyde in PBS.
  3. Rinse gently with Milli-Q® water. Repeat twice
  4. Rinse once with methanol and incubate in fresh methanol for 5 minutes.
  5. Aspirate the methanol. Add Wright’s stain to cover the inside membrane.
  6. Incubate for 30 seconds.
  7. Rinse gently with Milli-Q® water. Repeat three times.
  8. Observe as a wet mount.

Materials