| Oil
Red O Staining Procedure
Literature
Purpose:
This standard operating procedure (SOP) establishes the standard
for staining fat using the Oil Red O staining procedure.
Reagents
and Materials
Reagents
Oil Red O, Stock
Oil Red O 0.4gm
Isopropyl Alcohol 200ml
30% Isopropyl Alcohol
Isopropyl Alcohol 15ml
Distilled water 35ml
Oil Red O, Working
Use equal parts: Stock Oil Red O
30% Isopropyl Alcohol
Mix well and allow to stand 15 minutes (filter).
Materials
Tissue slides and control slides for special stain requested.
Microwave to bake slides.
Staining rack, 20 slide capacity.
Coverslips and glycerin jelly.
Hazards
and Precautions:
See Material Safety Data Sheets.
Sample Preparations:
Frozen tissue section slides cut at 8 microns.
Calibration
and Standardization:
A tissue section slide containing fat is stained with each procedure.
Rat adrenal is a good positive control tissue.
Procedure:
1.
Distilled water rinse.
2. 30% Isopropyl Alcohol.
3. Working Oil Red O - 15 minutes.
4. 30% Isopropyl Alcohol.
5. Distilled water rinse.
6. Harris Hematoxylin – 1 minute.
7. Distilled water rinse.
8. Mount in glycerin jelly.
Results:
Fat - red (Figure 1 arrows)
Nuclei - blue
Cherie
Chapman
James R. Turk
7/1/04
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