Masson’s Trichrome Stain Service: Reveal Fibrosis and Collagen with Precision 

At iHisto, we provide high-quality Masson’s Trichrome staining to clearly differentiate collagen, muscle fibers, and nuclei in tissue samples. Ideal for fibrosis scoring, liver pathology, and connective tissue analysis, our protocol ensures clean, consistent slides for both research and preclinical applications.

🧪 Overview 

Masson’s Trichrome is a three-color histological stain used to highlight:

• Collagen fibers – blue or green

• Muscle fibers and cytoplasm – red

• Cell nuclei – black or dark purple

This vivid contrast allows clear evaluation of fibrosis, scarring, and tissue remodeling — making it a cornerstone in toxicology, oncology, and pathology studies.

⚙️ How the Stain Works 

Masson’s Trichrome involves sequential dye application:

• Weigert’s Iron Hematoxylin – stains nuclei black

• Biebrich Scarlet–Acid Fuchsin – stains cytoplasm and muscle red

• Phosphotungstic/Phosphomolybdic Acid – removes red from collagen for cleaner separation

• Aniline Blue or Light Green – stains collagen blue or green

This combination delivers excellent separation of structural components, allowing detailed tissue evaluation.

🧬 Step-by-Step Staining Process

1. Fixation and Sectioning
   Formalin-fixed, paraffin-embedded tissues are sectioned onto slides.

2. Deparaffinization and Hydration
   Slides are processed through xylene and graded alcohols into water.

3. Nuclear Staining
   Weigert’s hematoxylin stains nuclei black.

4. Cytoplasmic Staining
   Biebrich Scarlet–Acid Fuchsin stains cytoplasm and muscle red.

5. Differentiation
   Phosphotungstic and/or phosphomolybdic acid removes excess red from collagen.

6. Collagen Staining
   Aniline Blue or Light Green selectively stains collagen fibers.

7. Dehydration and Mounting
   Slides are cleared and sealed for microscopy and digital scanning.

⚙️ Our Optimized Workflow

To ensure reproducibility and minimize human error, we use semi-automated equipment:

• Leica ST5020 Autostainer – consistent dye timing and control

• TS5025 Transfer Station – reduces processing variability

• CV5030 Coverslipper – ensures clean, bubble-free slide sealing

Whether for small batch academic studies or large-scale GLP programs, our system ensures consistent staining quality.

🔬 Applications of Masson’s Trichrome Staining 

Common use cases include:

• Liver fibrosis evaluation (NASH, cirrhosis – Ishak or METAVIR scoring)

• Cardiac fibrosis after infarction

• Renal pathology – glomerulosclerosis and interstitial fibrosis

• Tumor stroma analysis

• Skeletal or smooth muscle injury

• Dermal collagen mapping in skin research

This stain remains indispensable for fibrosis modeling and structural tissue analysis.

✅ Why Choose iHisto 

• ✅ Crisp differentiation of collagen, muscle, and nuclei

• ✅ Works with human, rodent, and primate FFPE tissues

• ✅ Compatible with standard and large-format slides

• ✅ Includes high-resolution whole-slide scanning

• ✅ Optional semi-quantitative fibrosis scoring and pathologist review

We support both exploratory and regulatory-stage studies with consistent slide quality and expert consultation.

❓ FAQs 

• What does Masson’s Trichrome stain detect?
  It stains collagen (blue/green), muscle (red), and nuclei (black) to assess fibrosis and tissue remodeling.

• Is this used in liver fibrosis models?
  Yes — it’s routinely used in liver research, particularly for fibrosis scoring systems like METAVIR or Ishak.

• Can I get digital slides?
  Absolutely. We provide whole-slide scanning and cloud or encrypted delivery.

• Do you offer scoring or quantification?
  Yes. We offer semi-quantitative fibrosis scoring or can work with your pathologist team for blinded interpretation.

📩 Request a Quote or Consultation 

Whether you're modeling fibrosis, tracking drug effects, or analyzing tissue remodeling, iHisto delivers Masson’s Trichrome staining with clarity, consistency, and confidence.

👉 Request a Quote or email info@ihisto.io