UCSF

Oil red O staining

Edited by Lucy 05/23/12

Materials

  1. Propylene glycol (kit included)
  2. Oil red o stain (kit included)
  3. Modified Mayer’s Hematoxylin (kit included)
  4. 10% Formalin
  5. 85% Propylene glycol

OIL RED O STAIN KIT : American MasterTech item#KTORO

Procedure

  1. Place frozen sections at RT 30min.
  2. Preheat Oil Red O Stain to 60°C.
  3. Place frozen sections in 10% Formalin for 2-5 minutes then rinse slide in tap water.(fixed slide may skip this step)
  4. Circle the tissue by Dako Pen.
  5. Place slide in Propylene glycol for 2 minutes.
  6. Place slide in Oil Red O Stain heated to 60°C for 6 minutes.
  7. Differentiate tissue in 85% propylene glycol solution for 1 minute.
  8. Rinse slide through 2 changes of distilled water
  9. Place slide in Modified Mayer’s Hematoxylin for 1 minute
  10. Rinse slide through 2 changes of tap water
  11. Rinse slide through 2 changes of distilled water
  12. Coverslip using an aqueous mounting medium

Results

  • Fat cells and neutral fat: RED
  • Nuclei: BLUE