Fat - red (Figure 1 arrows)
Nuclei - blue
Figure 1.
Oil Red O Staining Procedure for Lipids
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.
Positive
control: dog spleen with hemosiderosis
Results:
Fat - red (Figure 1 arrows)
Nuclei - blue
Figure 1.
Effective:
July 1, 2004
Cherie Chapman
James R. Turk
References:
Sheehan DC, Hrapchak BR: Theory and Practice of Histotechnology, 2nd Ed, CV Mosby, St. Louis, 1980, pp. 157.
Armed Forces Institute of Pathology Laboratory Methods in Histotechnology, eds. Prophet EB, Mills B, Arrington JB, Sobin LH, American Registry of pathology, Washington D.C. , 1992, pp. 157.