Martius Yellow-Acid Fuchsin-Aniline Blue Stain

 

 

Intended use:

Martius yellow-Acid fuchsin-Aniline blue stain is intended for fibrin staining, demonstrating the presence of fibrin in tissues and vascular cavity.

 

Principle:

It is based on multi-stain (Martius yellow-Scarlet blue, MSB) of Lendrum, which uses multiple anion dyes to mix or function successively. As the permeability varies among different tissues, using multiple anionic stains, each with a different molecular weight, allows the visualization of various tissue components. Briefly, compact erythrocytes are selectively stained in Martius yellow of low molecular weight, while fibrin and muscle fibers are stained red in acid fuchsin of medium molecular weight, and collagen fibers with loose structure are stained blue in aniline blue solution of high molecular weight.

 

Methods:

1. Fix tissues in Formalin/Mercuric chloride overnight and rinse in running tap water for one night. Dehydrate in typical manner and embed.

2. Deparaffinize slides in deionized water and remove mercury.

3. Place in Lazuline stain for 2~3 minutes and rinse briefly with distilled water.

4. Place in Mayer Hematoxylin Solution for 2~3 minutes and rinse briefly with distilled water.

5. Differentiate with 1% hydrochloric acid ethanol and wash in running tap water for 10 minutes. Wash in 95% ethanol.

6. Place in Martius yellow for 2 minutes. Rinse briefly with distilled water. Stain in Acid Fucshin for 10 minutes. Rinse briefly with distilled water.

7. Place in 1% Phosphotungstic acid for 5 minutes. Rinse briefly with distilled water.

8. Place in Aniline Blue Solution for 5~10 minutes. Rinse with 1% aqueous acetic acid to remove excess stain and differentiate.

9. Remove water around with filter paper. Dehydrate through absolute alcohol.

10. Clear in xylene and mount with mounting media.

 

Specifications:

 

Contents	6vialsx20ml/kit	6Btlsx100ml/kit	Components
Aniline Blue Solution	20ml	100ml	Aniline Blue
Mayer  Hematoxylin Solution	20ml	100ml	Hematoxylin
Martius yellow Solution	20ml	100ml	Martius yellow
Phosphotungstic acid	20ml	100ml	Phosphomolybdic Acid
Acid fuchsin	20ml	100ml	Acid fuchsin
Lazuline stain	20ml	100ml	Lazuline

Precaution:

1. Formalin/Mercuric chloride fixation is preferred, better than 10% formalin fixation. If fixed with Mercuric chloride, mercury must be removed and rinse thoroughly.

2. With this method, fresh fibrin will be stained red while old fibrin will be stained blue, which should not be confused with collagen fibers.

3. If necessary, observe microscopically after 1% Phosphotungstic acid processing until collagen fibers is colorless.

4. Tissue should be 4-5 µm thick.

 

Expected Results:

 

Fibrin	——	Vermeil
Muscle fibers	——	Red
Nuclei	——	Blue-brown
Old Fibrin	——	Indigo
Collagen fibers	——	Blue
Erythrocytes	——	Yellow