VitroView™ Masson’s Trichrome Stain Kit

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 Introduction Viedeo

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 2-3 seconds in distilled water.
11. Acetic Acid Solution for 2-3 seconds, 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 once for 2-3 seconds in distilled water.
12. 1% Acetic acid for 2-3 seconds , discard solution.
13. Dehydrate with 2 dip of 95% Ethanol and 2 changes of 100% Ethanol (2 minutes 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

More Images

Masson Trichrome Staining, Rat Heart	Masson Trichrome Staining, Mouse Lung
	Masson Trichrome Staining, Mouse Esophagus

Masson’s Trichrome Stain Kit Comparison Among Different Vendors

Vendor / Catalog#	VitroVivo Biotech / VB-3016	Abcam / ab150686	Millipore-Sigma / HT15
Components	1. Bouin’s Solution (1 x 250 mL) 2. Weigert’s Hematoxylin Solution A (1 x 125 mL) 3. Weigert’s Hematoxylin Solution B (1 x 125 mL) 4. Biebrich Scarlet-Acid Fuchsin Solution (1 x 250 mL) 5. Phosphomolybdic-Phosphotungstic Acid Solution (1 x 250 mL) 6. Aniline Blue Solution (1 x 250 mL) 7. 1% Acetic Acid Solution (1 x 250 mL)	1. 1% Acetic Acid Solution (1 x 125 mL) 2. Aniline Blue Solution (1 x 125 mL) 3. Biebrich Scarlet/Acid Fuchsin Solution (1 x 125 mL) 4. Bouin’s Fluid (1 x 125 mL) 5. Phosphomolybdic/Phosphotungstic Acid Solution (1 x 125 mL) 6. Weigert’s Iron Hematoxylin A (1 x 125 mL) 7. Weigert’s Iron Hematoxylin B (1 x 125 mL)	1. Biebrich Scarlet-Acid Fuchsin Solution (Cat. No. HT151-250ML) 2. Phosphotungstic Acid Solution (Cat. No. HT152-250ML) 3. Phosphomolybdic Acid Solution (Cat. No. HT153-250ML) 4. Aniline Blue Solution (Cat. No. HT154-250ML)
Website	https://vitrovivo.com/product/massons-trichrome-stain-kit/	https://www.abcam.com/en-us/products/assay-kits/trichrome-stain-kit-connective-tissue-stain-ab150686	https://www.sigmaaldrich.com/US/en/product/sigma/ht15#product-documentation
List Price (USD)	$369 / kit	$505 / kit	$429 / kit
Tests per Kit (1 mL/slide)	250 slides	125 slides	250 slides
Cost per Slide (1 mL/slide)	$1.48 / slide	$4.04 / slide	$1.72 / slide, plus cost of other reagents
Quality/Staining Images

Peer-reviewed publications and preprints that have cited the use of the VitroView™ Masson’s Trichrome Stain Kit (VB‑3016) kit

1. Ko KW, et al. (2021) — Integrated bioactive scaffold with polydeoxyribonucleotide and stem-cell-derived extracellular vesicles for kidney regeneration.
   ACS Biomaterials Science & Engineering 15(4):7575–7585.
   Uses VB-3016 Masson’s Trichrome to assess collagen and connective tissue in kidney tissue. VitroVivo

2. Park SY, et al. (2024) — Multifunctional vitamin D-incorporated PLGA scaffold with BMP/VEGF-overexpressed tonsil-derived MSC via CRISPR/Cas9 for bone tissue regeneration.
   Materials Today Bio 28:101254.
   Trichrome staining with VB-3016 was employed to evaluate collagen deposition during bone regeneration. VitroVivo

3. Lee JK, et al. (2024) — Osteoporotic bone regeneration via plenished biomimetic PLGA scaffold with sequential release system.
   Small 20:2310734.
   VB-3016 was used to visualize connective tissue and collagen in remodeled bone sections. VitroVivo

4. Barkovskaya A, et al. (2025) — Inhibition of PIKfyve kinase induces senescent cell death…, mouse model of idiopathic pulmonary fibrosis.
   bioRxiv preprint (2025.03.19.644224).
   Masson’s Trichrome with VB-3016 was applied to examine fibrosis and collagen remodeling in lung tissue. VitroVivo

5. Nam SH, et al. (2024) — Glycinamide facilitates nanocomplex formation… promotes osteoblast differentiation in vivo.
   Biomaterials Advances 10:947-959.
   Collagen deposition in bone and connective tissues was evaluated with the VitroView trichrome kit. VitroVivo

6. Bui HTD, et al. (2023) — Korean amberjack skin-inspired hyaluronic acid bioink for reconstruction of human skin.
   ACS Omega 8(25):22752-22761.
   VB-3016 was used to highlight collagen and connective tissue in engineered skin constructs. VitroVivo

7. Kim DS, et al. (2022) — Controlled vitamin D delivery with injectable hyaluronic acid-based hydrogel for restoration of tendinopathy.
   Journal of Tissue Engineering 13:1177/20417314221122089.
   Trichrome was used to assess collagen fiber organization in tendon repair. VitroVivo

8. Kim DS, et al. (2021) — Advanced PLGA hybrid scaffold with a bioactive PDRN/BMP2 nanocomplex for angiogenesis and bone regeneration…
   Science Advances 7:eabj1083.
   Collagen deposition and tissue integration were evaluated with VB-3016 trichrome staining. VitroVivo

9. Baek SW, et al. (2023) — Continuous NO dual-generation by ZnO nanoparticle conjugated with α-lipoic acid for functional biodegradable vascular stent.
   Chemical Engineering Journal 470:144174.
   Masson’s Trichrome was used for connective tissue and collagen analysis in vascular tissues. VitroVivo

10. Lee S, et al. (2024) — Injectable microparticle-containing hydrogel with controlled release of bioactive molecules for facial rejuvenation.
    Materials Today Bio 24:100890.
    Collagen and extracellular matrix remodeling were visualized using VB-3016. VitroVivo

11. Meca-Laguna G, et al. (2025) — γδ T Cells Target and Ablate Senescent Cells in Aging and Alleviate Pulmonary Fibrosis.
    bioRxiv preprint (2025.05.05.652251).
    Trichrome staining with VB-3016 was used to evaluate collagen deposition in lung fibrosis. VitroVivo

12. Hypertension-induced heart failure disrupts cardiac sympathetic innervation — PMC Article.
    This paper reports using the VitroView Masson’s Trichrome Stain Kit (VB-3016) to perform trichrome staining on cryosections of cardiac tissue. PMC

Why Choose the VitroView™ Masson’s Trichrome Stain Kit for Your Research?

1. Cost-effective
2. Easy to use
3. High quality with strict quality control and assurance
4. Validated and trusted by numerous peer-reviewed publications

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