Mucicarmine Staining Principle and Procedure

Mucicarmine staining principle and procedure post we have briefly explained about Mucicarmine staining method’s principle, requirements, mucicarmine staining procedure, result with Label.

Mucicarmine Staining Principle and Procedure

To stain mucin which is a secretion produced by a variety of epithelial cells and connective tissue cells. Epithelial cells secrete too much mucin in certain inflammations and intestinal carcinomas. 

Finding mucin positive tumour cells in an area that does not contain mucin generating cells indicates that the tumour did not emerge from that location, which is why the mucicarmine technique is beneficial in pinpointing the site of a primary tumour. It can also be used to stain encapsulated fungi, such as cryptococcus.

Principle

Carmine-aluminium complexes that are positively charged form bonds with acid mucins that are negatively charged. The carmine solution is extremely complicated, and the complete explanation is unknown. 

Heavily sulphated mucins react differently, with neutral mucins not staining at all and acidic mucins (especially hyaluronic acid) staining strongly. 

So as not to mask the staining result, counterstains which also demonstrate mucins (eg Ehrlich’s haematoxylin) are to be avoided.

Requirements

Chemicals

1. Hematoxylin Crystals

2. Alcohol

3. Ferric Chloride

4. Hydrochloric Acid

5. Carmine

6. Aluminium Hydroxide          

7. Aluminum Chloride Anhydrous

Equipment’s

1. Light Microscope

2. Stirring Hot Plate

3. Magnetic Stir Bars

4. Coplin Jars

5. Microwave Oven

Reagents

1. Alum haematoxylin

Solution A: Hematoxylin crystals: 2.0 gm, Alcohol, 90%: 100.0 ml. Solution B: Ferric chloride: 4.0 ml, Distilled water:  95.0 ml, Hydrochloric acid, concentrated: 1.0 ml. (Note: For use mix equal parts of each solution).

2. Mucicarmine solution

Carmine: 1.0 g, 50% alcohol: 100.0 ml, Aluminium hydroxide: 1.0 g, Aluminum Chloride Anhydrous: 0.5 g. To make the carmine, grind it to a fine powder, then pour it in a 500 mL Ehrlenmeyer flask with the 50% alcohol and mix well. Mix in the aluminium hydroxide. Mix in the aluminium chloride. Bring to a boil, and then reduce to a low heat for two to three minutes. Allow to cool before filtering into a reagent bottle. Refrigerate mucicarmine staining reagent at 4°C .

Procedure

1. Submerge the slides in distilled water.

2. For 2 minutes, stain the nuclei with alum haematoxylin.

3. Rinse under running water

4. Differentiate in acid alcohol

5. Rinse under running water

6. Blue in Scott’s tap water substitute

7. Rinse under running water

8. 20 minutes staining with mucicarmine solution

9. Rinse under running water

10. Dehydrate, clear, and mount your work.

Result

Southgate's Mucicarmine Staining

Mucins: Magenta

Nuclei: Blue

Cryptococcus neoformans: Magenta

Further Readings

Reference