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