Oil Red O Stain

Oil Red O Stain

Assay-kits

Product code: ACMBS-24
Specifications: 2x20mL, 2x250mL (Box)
Intended use: Used for staining of fat in tissue cells
Store temperature: 5℃～30℃
Expiry date: 18 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationPrecautions

Principle

Oil Red O, also known as Sudan Red 5B. The principle of staining lipids is believed to the physical properties of solution or adsorption by effect of solution. Dissolved Sudan colorants in organic solvents. Dyes exhibit a greater solubility in lipoid substances of frozen tissue than that in original solvents. So, during staining, dyes will migrate from organic solvents, to lipids, that staining fat in tissue cells.

Main components

Reagent composition	Main ingredient
1. Oil Red O stock solution	Oil Red O
2. Hematoxylin solution (Mayer)	Hematoxylin

Sample requirement

Frozen section or carbon wax section, thickness of about (6～8) μm.

Test procedure

1. Rinse tissue in 70% ethanol for 5 seconds.
2. Prepare Oil Red O working solution: (6 ml of Oil Red O stock solution and 4 ml of deionized water).
3. Mix Oil Red O working solution well and stand for 10～20 minutes. Try to absorb the upper liquid and stain with the upper liquid for 5～10 minutes.
4. Rinse in 70% ethanol for several seconds to remove excess stain. Rinse in distilled water for 30 seconds.
5. Stain with hematoxylin solution (Mayer) for 2 minutes. Rinse in running water for 5 minutes.
6. Remove excess water around by filter paper.
7. Mount with Arabic gum or glycerin gelatin.

Resreult interptation

Neutral Fat	Dark orange-red
Nuclei	Blue

Precautions

1. After the frozen section is prepared, it can be stained without fixing again, it can also be fixed in 10% formaldehyde solution for 1 to 2 minutes, and the fixation time should not be too long to avoid lipid dissolution.
2. Since fats are readily soluble in organic solvents, it is generally not shown that fats can be treated like paraffin sections, but rather displayed by frozen section staining.
3. The frozen tissue used for lipid staining should not be too thin. Otherwise, the lipid content may be lost.
4. After the Oil red O working solution is prepared, it should be dyed after standing for 10 to 20 minutes. If the standing time is too long, it may cause light dyeing. Precipitated particles will precipitate and sink to the bottom of the container, so try to use the upper working solution, to avoid precipitation of the particles.
5. The counterstaining time in hematoxylin solution (Mayer) should not be too long.
6. The tissue staining will not last for too long, so observe and take pictures as soon as possible.
7. This reagent should be used by professionals and interpretation of the results. Read the instruction manual carefully before use, use it within the validity period, and do personal hygiene protection.
8. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
9. The production date, production batch number and expiration date are shown in the outer packaging.

References

1. Chinese Medical Association. Clinical technical operating specifications-pathology volumes [M]. People's Military Medical Publishing House.

2. Ling Qibo. Practical pathology special staining and histochemistry technology [M]. Guangdong Higher Education Press.

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