Dear Saber, here some protocols and notes that could be useful
Neutral mucins may be stained red, brownish red, and violet, respectively, by carmine, hematoxylin, and orcein from appropriate alkaline solutions. Carmine and hematoxylin in concentrations of 0.2-1% are dissolved in 60-70% alcohol containing 1% potassium carbonate; staining times are 15 to 30 minutes. The stained sections are rinsed in 95% or absolute alcohol prior to xylene and mounting. The staining of these mucins is blocked by mild bromine oxidation. By using alcian blue 0.1% in 3% acetic acid for 5 minutes prior to the above stains, mucins may be characterized in the same preparation as acid, neutral or mixed.
Mucicarmine: Staining Protocol
Sample Type Required: Deparaffinized and re-hydrated tissue sections on positively charged slides. Fixative: 10% Neutral Buffered Formalin Step Reagent Time Technical Notes1) Weigert's Iron Hematoxylin Working Solution½ Solution A (Hematoxylin and 95% Alcohol) and ½ Solution B (Distilled Water, Hydrochloric Acid and 29% Ferric Chloride Solution), 7 Minutes. Be careful not to overstain with Hematoxylin as this can obscure the carminophilic tissue elements. If Gills hematoxylin is used in place of Weigert's Iron Hematoxylin the mucin may have a bluish cast.
2) Running Tap Water 10 Minutes. 3) Working Mucicarmine Solution: 10mL Mucicarmine Stock Solution and 40mL Distilled Water, 60 Minutes. Stock Mucicarmine should be stored refrigerated to slow down deterioration of the solution. Expiration of the solution should be closely monitored. Do not freeze solution to avoid frequent freeze-thaw cycles that may cause deterioration. 4) Distilled Water, 1-2 Changes. 5) Metanil Yellow 0.25% Solution 30 Seconds to 1 Minute. Be careful not to overstain with Metanil Yellow as this can obscure carminophilic tissue elements. Tartrazine can be used in place of Metanil Yellow.
Expected ResultsMucin = Deep Rose Red Capsule of Cryptococcus = Deep Rose Red Nuclei = Black Other tissue elements = Blue or yellow. Post Staining Procedure: Tissue sections should be rinsed well in distilled water, dehydrated with 95% and absolute alcohols, cleared and cover-slipped. Mucicarmine stock solution should be stored refrigerated Room temperature storage will accelerate the chemical breakdown of the solution. Frozen storage is not recommended to avoid repeated freeze-thaw cycles that may also affect the chemical reactivity of the stain.
Good luck!
I have used Tartrazine as a counterstaining because the particular method I was using at he time called for it, but the colour of mucins should be clearer without it.
Matthew is right that using celestine blue (which is a mordant dye) after Mayer's will help to have good nuclear stain contained to nuclei. I have used this method on some very damaged forensic research samples and it worked very well.
Barbara's exceptionally detailed description of the method deserves a prize. reading it I almost expected additional note on preparation of Mucicarmine stock solution from scratch, but it can be bought (e.g. from Sigma), so of course would be unnecessary.
Good luck with your study
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