Sample Type Required: Deparaffinized and re-hydrated tissue sections on positively charged slides.
Fixative: 10% Neutral Buffered Formalin
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 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 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% 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.
