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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.

 

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Luxol Fast Blue Stain Variations

We did this as a test to see different versions of luxol fast blue (LFB). LFB is a stain for the myelin sheath or myelinated axons in brain and spinal cord.  We tested on rat brain.  There are 4 variations.  They are; LFB / Cresyl violet (CR), LFB /H&E, LFB / Periodic acid schiff (PAS) and LFB / nuclear fast red (NFR).   The H&E and NFR versions are for background staining only.  The PAS could show glycogen or fungus present and the CR shows nissl bodies.

Rat brain LFB / CV 20x
Rat brain LFB / CV 20x
Rat brain LFB / H&E 20x
Rat brain LFB / H&E 20x
Rat brain LFB / NFR 20x
Rat brain LFB / NFR 20x
Rat brain LFB / PAS 20x
Rat brain LFB / PAS 20x