VitroView™ Masson’s Trichrome Stain Kit

$299.00

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SKU: VB-3016 Category:

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:

  1. Deparaffinize in Xylene I for 6 minutes and II for 6 minutes.
  2. 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.
  3. Prepare Weigert’s working hematoxylin: mix Weigert’s Hematoxylin Solution A and B at 1:1 ratio.
  4. Weigert’s working hematoxylin for 10 minutes. (Note: Discard after use.)
  5. Blue in running tap water for 5 minutes, rinse once for 1 min in distilled water.
  6. Biebrich Scarlet-Acid Fuchsin Solution for 5 minutes. (Note: Solution may be used twice only, then discarded.)
  7. Rinse once for 1 min in distilled water. 
  8. Phosphotungstic/phosphomolybdic acid solution for 10 minutes, discard solution.
  9. Transfer directly into Aniline Blue Solution for 10 minutes.
  10. Rinse once for 1 min in distilled water.
  11. Acetic Acid Solution for 1 minute, discards solution.
  12. Dehydrate with 2 changes of 95% Ethanol and 2 changes of 100% Ethanol (2 minute per change).
  13. Clear with 3 changes of xylene (5 minutes per change) and coverslip with Permount or other suitable organic mounting medium.

For Frozen Sections:

  1. Fix frozen sections in 10% formalin (not Zinc) for 30 minutes.
  2. Rinses in distilled water 3×3min.
  3. Mordant in Bouin’s Solution, 60°C for 1 hour.
  4. Wash in running tap water for 5 minutes to remove the picric acid.
  5. Weigert’s working hematoxylin for 7 minutes. (Note: Discard after use.)
  6. Blue in running tap water for 5 minutes, rinse in distilled water.
  7. Biebrich Scarlet-Acid Fuchsin Solution for 5 minutes. (Note: Solution may be used twice only, then discarded.)
  8. Rinse in distilled water. 
  9. Phosphotungstic/phosphomolybdic Acid Solution for 10 minutes, discard solution.
  10. Transfer directly into Aniline Blue Solution for 5 minutes.
  11. Rinse in distilled water.
  12. 1% Acetic acid for 1 minute, discard solution.
  13. Dehydrate with 2 changes of 95% Ethanol and 2 changes of 100% Ethanol (2 minute per change).
  14. 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

Note: This Trichrome Stain Kit is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Precautions: Handle with care. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.

User Manual and Material Safety Data Sheet (MSDS)  (PDF)

VB-3016 User manual

VB-3016 MSDS

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