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Updated: Jun 17, 2026
11:39
Using Unfixed, Frozen Tissues to Study Natural Mucin Distribution
Published on: September 21, 2012
De-staining and re-staining mucins in formalin fixed paraffin sections
1Barry University School of Podiatric Medicine, Miami Shores, Florida 33161, USA.
Summary
Histological sections can be de-stained and re-stained using simple protocols. This allows for the re-use of valuable tissue samples, improving research efficiency.
Area of Science:
- Histopathology
- Tissue Staining Techniques
Background:
- Re-staining histological sections is often necessary.
- Effective de-staining protocols are crucial for successful re-staining.
- Formalin-fixed paraffin-embedded (FFPE) tissues are commonly used.
Purpose of the Study:
- To describe simple and effective de-staining protocols for various histological stains.
- To enable re-staining of previously stained FFPE tissue sections.
Main Methods:
- De-staining protocols were developed for specific stains including Mucihematoxylin, Mucicarmine, Zirconyl hematoxylin, Alcian blue, Colloidal iron, and Periodic acid-Schiff (PAS).
- Specific chemical agents were used for de-staining, such as acid alcohol, periodic acid, Sinha's fixative, trifluoroacetic acid, household bleach, hydrogen peroxide, and ammonium hydroxide.
- The efficacy of de-staining was assessed by the ability to subsequently re-stain sections.
Main Results:
- Mucihematoxylin and Mucicarmine stains were effectively removed using acid alcohol.
- Zirconyl hematoxylin was removed with periodic acid or Sinha's fixative.
- Alcian blue was removed with 5% trifluoroacetic acid in dichloromethane.
- Colloidal iron was bleached using 1% household bleach in alcohol.
- PAS stains were removed with hydrogen peroxide or ammonium hydroxide.
- Most de-stained sections could be successfully re-stained with Mucihematoxylin, PAS, or Gabe's trichrome.
Conclusions:
- Simple and effective de-staining protocols exist for common histological stains.
- These methods facilitate the re-staining of FFPE tissue sections, preserving valuable samples.
- The described protocols enhance the utility of existing histological preparations for further analysis.
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