Oil Red O Stain Protocol

This is one of the stains done on frozen tissues for lipids or fat globules.

Safety equipment: Work under a hood with lab coat, gloves, and glasses.
Use on frozen sections only

1. Hydrate slides in running water———————– 10 minutes
2. Stain in working Oil Red O ————————— 10 minutes
3. Rinse in running water ——————————- 2 minutes
4. Rinse in 60% isopropanol —————————– 5 minutes
5. Rinse in running water ——————————- 2 minutes
6. Counterstain with Harris Hematoxylin —————– 20 seconds
7. Blue in running water ——————————– 5 minutes
8. Coverslip with glycerin jelly or aqueous mounting media

Results:
Fat Globules – Orange Red
Nuclei – Blue

Notes:
• For tissues that do not stay on slides, do not rinse slides in water prior to staining.
• Dextrin must be made for new working solution.
• Dextrin is only good for 1 month.

 

An Expert Histology Service


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