List of Stains
Types of Stains
Mechanisms of Staining
- Ionic bonding
- Hydrogen bonding
- Histochemical
- Use a non-coloured substance to produce a coloured, non-diffusable end-product in the tissue
- Silver Staining
- Argentaffin metallic impregnation
- Argyophillic metallic impregnation
Summary of Conventional Staining Techniques
Stain | Uses | Tissue Components Targeted | Mechanism of Staining (Primary Stain) | Stains Used
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Other Reagents | Controls |
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Hematoxylin & Eosin (H&E) |
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Periodic Acid-Schiff (PAS) |
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Alcian Blue (AB) |
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Ionic bonding | |||
Alcian Blue + Periodic Acid-Schiff (AB PAS) | Identifying multiple different types of carbohydrate groups | |||||
Congo Red |
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Hydrogen bonding | ||||
Masson Trichrome |
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Connective Tissue Stain
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Porosity and ionic bonding | No primary stain.
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Gordon and Sweet's Reticulin |
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Connective Tissue Stain
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Metallic impregnation
Counterstain
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Verhoeff's Van Giesen | Connective Tissue Stain
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Connective Tissue Stain
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Van der Waal's
Porosity and ionic bonding
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Oil Red O | Connective Tissue Stain
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Selective solubility | |||
Gram | Differential Microorganism Stain
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Microorganism Stain
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Ionic bonding |
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Positive: appendix, any tissues containing bacteria |
Ziehl-Neelsen | Differential Microorganism Stain
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Microorganism Stain
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Ionic bonding |
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Positive: any tissue with acid fast bacteria (e.g., M. tuberculosis) |
Grocott Methenamine Silver | Microorganism Stain
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Microorganism Stain
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Argentaffin metallic impregnation
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Positive: any tissue with fungi |
Perl's Prussian Blue | Iron Pigment Stain
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Histochemical |
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Positive: spleen, bone marrow, liver | |
Masson Fontana | Pigment Stain
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Argentaffin metallic impregnation |
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Positive: skin (melanin), small intestine (argentaffin granules)
Need 4 slides to confirm Melanin!
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Von Kossa | Pigment Stain for Calcium
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Metallic substitution - silver impregnation |
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Positive: any calcified tissue (e.g., bone)
Can be used as negative control for:
Need 4 slides for Ca neg control
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Jones Methenamine Silver | Connective Tissue Stain
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Argentaffin metallic impregnation |
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Positive: normal kidney |
Toluidine Blue | Connective Tissue Stain
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Metachromasia and hydrogen bonding |
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Positive: tissue with mast cells (skin, small intestines) |
Summary of Silver Staining Methods
Silver staining can be done via two methods: argentaffin or argyophillic staining.
- Argentaffin staining has fewer steps as the tissue components can auto-reduce the silver and do not require sensitization.
- Argyophillic methods require a sensitizing and reducing agent.
Argentaffin Methods* | Argyophillic Methods | Metallic Substitution | |||
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Stain | Grocott's Methenamine Silver (GMS) | Jones Methenamine Silver (JMS) | Masson Fontana (MF) | Gordon & Sweet's Reticulin (GSR) | Von Kossa (VK) |
Oxidation
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Chromic acid
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Periodic acid
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Gram's iodine
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Bleaching
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Sodium bisulphate | Sodium thiosulphate | |||
Sensitization
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Impregnation
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Methenamine silver | Methenamine silver | Fontana ammoniacal silver | Silver nitrate | |
Reduction
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Light or hydroquinone | ||||
Toning
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Gold chloride | Gold chloride | Gold chloride | ||
Fixation
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Sodium thiosulphate (hypo) | Sodium thiosulphate (hypo) | Sodium thiosulphate (hypo) | Sodium thiosulphate (hypo) | |
Counterstain
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LG or NFR | LG or NFR | LG or NFR | LG, NFR, or van Gieson |
- The bleaching step is only required when the reagents themselves are coloured or cause discolouration.
- Sensitization step is not required in argentaffin methods as silver can attach directly to the target tissue.
- Argentaffin methods involve auto-reducing tissue. The tissue reduces the silver itself, so no reduction agents are needed.
Summary of Immunohistochemistry Techniques
Technique | |||
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Gram Stain
- Staining technique: Differential staining (ionic bonding to cell wall)
- Primary Stain: Crystal violet
- Targets: Gram positive bacteria (purple/blue)
- Counterstain: Safranin
- Targets: Gram negative bacteria (pink/red)
Ziehl-Neelsen
- Staining technique: Differential staining/acid fastness (ionic bonding)
- Primary Stain:
- Targets: Acid-fast bacteria (e.g., Mycobacteria)
- Counterstain:
- Targets:
Grocott Methenamine Silver
- Staining technique:
- Primary Stain:
- Targets:
- Counterstain:
- Targets:
Perl's Prussian Blue
- Staining technique:
- Primary Stain:
- Targets:
- Counterstain:
- Targets:
Masson Fontana
- Staining technique:
- Primary Stain:
- Targets:
- Counterstain:
- Targets:
Von Kossa
- Staining technique:
- Primary Stain:
- Targets:
- Counterstain:
- Targets:
Jones Methenamine Silver
- Staining technique:
- Primary Stain:
- Targets:
- Counterstain:
- Targets:
Toluidine Blue
- Staining technique:
- Primary Stain:
- Targets:
- Counterstain:
- Targets: