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Schiff’s Reagent Reactions

Staining Method

Schiff’s reagent reactions, also known as Schiff Tests, can be used to visualize aldehydes in a tissue sample.

Introduction

Schiff’s reagent, also known as fuchsin-sulphurous acid,  is a solution that will combine chemically with aldehydes to form a bright red product. Strictly speaking, ketones also react but for all practical purposes they can be ignored. Many tissue components can be stained this way. Schiff’s reagent is made from pararosanilin treated with sulphurous acid. This causes a disruption of the chromophore by the addition of a sulphonic acid group to the central carbon. The sulphurous acid is produced by dissolving sulphur dioxide in water.

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Periodic acid Schiff staining in liver biopsy of glycogen storage disorder, 20X magnification

periodic-acid-schiff-staining-liver-biopsy-glycogen-storage-disorder-600x400

Figure 1. Periodic Acid Schiff Staining in Liver Biopsy of Glycogen Storage Disorder

Positive periodic acid Schiff (PAS) staining (dark purple) due to intracytoplasmic accumulation of glycogen in a liver biopsy of glycogen storage disorder shows uniformly distended hepatocytes with clear to pale eosinophilic cytoplasm. 20X magnification. Liver in glycogen storage disease, PAS stain by Department of Pathology, Calicut Medical College is licensed under CC BY-SA 4.0.

Periodic acid Schiff staining of esophageal candidiasis

periodic-acid-schiff-staining-esophageal-candidiasis-600x400

Figure 2. Periodic Acid Schiff Staining of Esophageal Candidiasis

Periodic acid Schiff (PAS) staining (dark purple) demonstrates fungal morphology in esophageal candidiasis. Esophageal Candidiasis, PAS Stain by KGH is licensed under CC BY-SA 3.0.

 

Types of Reactions

Periodic Acid-Schiff (PAS) Reaction

The most common application for Schiff’s reagent is the Periodic Acid Schiff reaction for demonstrating carbohydrates in tissue sections.

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Nucleal Reaction

Given its sensitivity towards detecting aldehydes, Schiff’s reagent can be used to demonstrate deoxyribosenucleic acid (DNA), especially in contrast to unstained ribosenucleic acid (RNA).

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Plasmal Reaction

The plasmal reaction can be used to demonstrate plasmalogens to visualize myelin.

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Aldehyde-Schiff Reaction

By targeting amino groups, Schiff’s reagent can be used to demonstrate proteins in a tissue sample.

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Schiff’s Reagent Formulations

There are numerous variants or formulations of Schiff’s reagent that may be used for demonstrating aldehydes in samples. Find formulation details, including detailed steps for preparation, storage considerations, and reaction notes for over 10 variants below.

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Alternatives to Schiff’s Reagents

While reactions with Schiff’s reagent are the most common way of staining aldehydes, there are alternative methods. For example, a silver reduction reaction is a popular method for the demonstration of fungi and basement membranes. Other alternatives for demonstrating aldehydes include diazonium coupling, osmium reduction, or thiosemicarbazide reduction.

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Protocols

Find step-by-step instructions to complete a variety of Schiff’s reagent reactions, with detailed protocols for tissue preparation, staining, and supplementary procedures.

References

  1. Pearse, A. G. E., (1968, 1972)
    Histochemistry: Theoretical and Applied, Ed. 3
    Churchill Livingstone, Edinburgh, London, UK
  2. Kiernan. J.A., (1999)
    Histological and histochemical methods: Theory and practice, Ed. 3
    Butterworth Heinemann, Oxford, UK.
  3. Culling C.F.A., (1974)
    Handbook of histopathological and histochemical techniques Ed. 3
    Butterworth, London, UK.
    Citing:
    Kasten, F.H., (1958), Stain Technology, vol.33, pp.39
    Kasten, F.H., (1959), Histochemie, vol.1, pp.466
  4. Lillie, R.D., (1954)
    Histopathologic technique and practical histochemistry Ed.2
    Blakiston, New York, USA.