Mucicarmine: Staining Protocol

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Mucicarmine: Staining Protocol

Sample type required: Deparaffinized and re-hydrated tissue sections on positively charged slides.

Fixative: 10% NBF


Step

Reagent

Time

Technical Notes

1

Weigert's iron hematoxylin working solution:

½ 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:

10 mL mucicarmine stock solution and 40 mL 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 no to overstain with metanil yellow, as this can obscure carminophilic tissue elements.

Tartrazine can be used in place of metanil yellow.

Expected results:

  • Mucin = 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% alcohol, followed by 100% alcohols, cleared with xylene (or xylene substitute), and cover-slipped.

Procedural notes:

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.

Mucicarmine stain demonstraiting goblet cells containing mucin