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Why Do We Use Special Stains in Histology?

  • Writer: Eghosa Arovo
    Eghosa Arovo
  • Jun 12
  • 3 min read

While routine haematoxylin & eosin (H&E) staining provides excellent general morphology, many research questions demand more specific tissue components:


  • Visualising extracellular matrix and connective tissue

  • Detecting mineral deposits or pigments

  • Highlighting specific fibre types

  • Differentiating carbohydrates or lipids


Special stains employ targeted chemistries to bind unique biomolecules, giving you contrast and clarity that H&E alone cannot achieve. In cancer research and broader biomedical studies, they can reveal subtle changes in tumour stroma, fibrosis, iron overload, or basement membrane integrity—crucial clues for understanding disease mechanisms.


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Key Special Stains and Their Applications


1. Sirius Red


  • What it stains: Collagen fibres (types I and III)

  • Why use it: Under polarized light, Sirius Red–stained collagen shows brilliant birefringence, allowing quantitative assessment of fibrosis or extracellular matrix remodelling in tumours and surrounding stroma.

Sirius Red staininig on Intestine. Fibres stained with dark-red.
Sirius Red staininig on Intestine. Fibres stained with dark-red.

2. Perls’ Prussian Blue

  • What it stains: Ferric iron (Fe³⁺) deposits

  • Why use it: Ideal for detecting hemosiderin or iron overload in tissues—useful in studies of haemochromatosis models, tumour-associated macrophages, or post-therapy iron deposition.

Liver sample stained with Perl's Prussian Blue. Blue dots are Ferric iron deposits.
Liver sample stained with Perl's Prussian Blue. Blue dots are Ferric iron deposits.

3. Reticulin (Gomori’s Silver)

  • What it stains: Reticular (type III collagen) fibres

  • Why use it: Highlights the fine “reticular” network that supports cellular architecture—commonly used in lymphoma grading, tumour vascular pattern analysis, and bone marrow stromal assessment.

Liver sample stained with Reticulin. Reticulin fibres are dark-brown/black silhouettes, and background has been stained with Nuclear Fast Red.
Liver sample stained with Reticulin. Reticulin fibres are dark-brown/black silhouettes, and background has been stained with Nuclear Fast Red.

4. Masson’s Trichrome (MTC)

  • What it stains: Collagen (blue or green), muscle fibres (red), nuclei (black)

  • Why use it: Offers a colourful separation of muscle, collagen, and cellular components—widely applied in assessing tumour desmoplasia, muscle invasion, and fibrosis quantification.

Layers of Skin tissue stained with Masson's Trichrome (MTC).
Layers of Skin tissue stained with Masson's Trichrome (MTC).

5. Elastic Van Gieson (EVG)

  • What it stains: Elastic fibres (black), collagen (red), other tissue (yellow)

  • Why use it: Excellent for visualising vascular elastic lamina, basement membranes, and elastic fibre integrity—key in angiogenesis studies and vascular remodelling in tumours.

Blood vessel stained with EVG
Blood vessel stained with EVG

Integrating Special Stains into Your Research Workflow


  1. Define your question: Are you measuring fibrosis, tracking iron, or mapping vascular changes?

  2. Select appropriate stains: Combine special stains with H&E or immunohistochemistry for multiplex insights.

  3. Quantitative analysis: Use digital slide scanners and image analysis software to quantify stain-positive area or fibre thickness.

  4. Expert interpretation: Partner with histology specialists to ensure optimal staining protocols and accurate data interpretation.



Why LabNexus for Special Stains?

At LabNexus, we offer:

  • A panel of singleplex and multiplex special stains, validated for research use only

  • High-throughput slide scanning and digital analysis pipelines

  • Expert technical support, from protocol selection to data reporting


Whether you’re exploring tumour microenvironment, fibrosis pathways, or vascular remodelling, our tailored histology services empower you to generate reliable, reproducible data.


Click Here to submit your request or make an inquiry regarding your project.


References

  1. Bancroft JD, Gamble M, editors. Theory and Practice of Histological Techniques. 8th ed. Churchill Livingstone/Elsevier; 2020.

  2. Junqueira LC, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy for collagen visualization. Histochem J. 1979;11(4):447–455. doi:10.1007/BF01002772

  3. Puchtler H, Waldrop FS, Valentine LS. A study of the specificity of the reticulin stain. J Histochem Cytochem. 1965;13(4):275–285. doi:10.1177/13.4.275

  4. Pearse AGE. Histochemistry: Theoretical and Applied. 4th ed. Churchill Livingstone; 1985.

 
 
 

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