Martius Scarlet Blue (MSB) stain kit of seven reagents used for the visualization of fibrin, especially from older groups. This method is a modification of Masson Trichrome and is ideal for studying connective tissue and vascular pathology.
The kit contains:
- Yellow Martius Sat. Alcoholic with 2% Phosphotungstic Acid
- 1% phosphotungstic acid solution
- Heigerts’ Hematoxylin (Solution A)
- bright crystal scarlet
- Aniline blue (1% aqueous)
- Heigerts’ Hematoxylin (Solution B)
- Procedure time: 45 minutes (approximate)
- Shelf life: 3 years (from the date of manufacture)
- Storage: 15-25°C.
- Fibrin: Red (early fibrin may stain yellow and old fibrin blue)
- Muscle: pale red
- Cores: Blue/black
- Collagen; basement membranes; reticulin: Blue
- Elastic fibres: Blue
- Erythrocytes: Yellow
1. Prepare fresh Weigert’s Hematoxylin by mixing equal volumes of Solution A and B as required.
2. Deparaffinize sections, hydrate with alcohols and rinse with tap water
3. Stain nuclei with Weigert’s iron hematoxylin for 10 minutes.
4. Wash quickly with water and differentiate in 1% acid alcohol solution (RRSP187 – not supplied), leaving nuclei slightly overstained.
5. Wash well under running tap water then Blue in water or Scotts tap water
6. Wash well under running water
7. Rinse with 95% alcohol (not supplied)
8. Stain with Martius yellow solution for 5 minutes.
9. Wash quickly with running tap water
10. Stain with scarlet crystal solution for 5 minutes.
11. Wash with running tap water
12. Treat with a phosphotungstic acid solution for 10 minutes, and check microscopically. Treat for an additional 5 minutes if necessary.
13. Wash with running tap water
14. Stain with Aniline Blue Solution for 5 minutes.
15. Wash with tap water
16. Dehydrate, rinse and mount