VitroView Masson's Trichrome Stain Kit (#VB-3016)
Description
Masson’s Trichrome Stain Kit is used to differentiate between collagen and smooth muscle in tumors, and the increase of collagen in diseases such as cirrhosis. It is a routine stain for liver and kidney biopsies. This kit can be used for FFPE and frozen tissue slides.
Kit Contents
- VB-3016-1 Bouin’s solution———————————————————250 ml
- VB-3016-2 Weigert’s Hematoxylin Solution A——————————125 ml
- VB-3016-3 Weigert’s Hematoxylin Solution B——————————125 ml
- VB-3016-4 Biebrich Scarlet-Acid Fuchsin Solution———————–250 ml
- VB-3016-5 Phosphomolybdic-Phosphotungstic Acid Solution —250 ml
- VB-3016-6 Aniline Blue Solution————————————————-250 ml
- VB-3016-7 1% Acetic Acid Solution———————————————250 ml
Protocol
For formalin-fixed, paraffin-embedded (FFPE) tissue sections:
- Deparaffinize in Xylene I for 6 minutes and II for 6 minutes.
- Rehydrate: ethanol 100% (2 minutes)x2; ethanol 95% (2 minutes)x2; ethanol 70% (2 minutes)x1; Rinse in distilled water (5 minutes).Mordant in Bouin’s Solution, 60°C for 1 hour.
- Prepare Weigert’s working hematoxylin: mix Weigert’s Hematoxylin Solution A and B at 1:1 ratio.
- Weigert’s working hematoxylin for 10 minutes. (Note: Discard after use.)
- Blue in running tap water for 5 minutes, rinse once for 1 min in distilled water.
- Biebrich Scarlet-Acid Fuchsin Solution for 5 minutes. (Note: Solution may be used twice only, then discarded.)
- Rinse once for 1 min in distilled water.
- Phosphotungstic/phosphomolybdic acid solution for 10 minutes, discard solution.
- Transfer directly into Aniline Blue Solution for 10 minutes.
- Rinse once for 2-3 seconds in distilled water.
- Acetic Acid Solution for 2-3 seconds, discards solution.
- Dehydrate with 2 changes of 95% Ethanol and 2 changes of 100% Ethanol (2 minute per change).
- Clear with 3 changes of xylene (5 minutes per change) and coverslip with Permount or other suitable organic mounting medium.
For Frozen Sections:
- Fix frozen sections in 10% formalin (not Zinc) for 30 minutes.
- Rinses in distilled water 3×3min.
- Mordant in Bouin’s Solution, 60°C for 1 hour.
- Wash in running tap water for 5 minutes to remove the picric acid.
- Weigert’s working hematoxylin for 7 minutes. (Note: Discard after use.)
- Blue in running tap water for 5 minutes, rinse in distilled water.
- Biebrich Scarlet-Acid Fuchsin Solution for 5 minutes. (Note: Solution may be used twice only, then discarded.)
- Rinse in distilled water.
- Phosphotungstic/phosphomolybdic Acid Solution for 10 minutes, discard solution.
- Transfer directly into Aniline Blue Solution for 5 minutes.
- Rinse once for 2-3 seconds in distilled water.
- 1% Acetic acid for 2-3 seconds , discard solution.
- Dehydrate with 2 dip of 95% Ethanol and 2 changes of 100% Ethanol (2 minutes per change).
- Clear with 3 changes of xylene (5 minutes per change) and coverslip with Permount or other suitable organic mounting medium.
Expected results
- Cytoplasm, keratin, muscle fibers, Erythrocytes ——— red
- Nuclei ———————————————————————— black
- Collagen and mucus ————————————————- blue
VitroView Masson's Trichrome Stain Kit (#VB-3016)
SKU: VB-3016