Mucicarmine Stock Stain, Mayer

SOLUTION:

Additionally Needed For Mucin, Mayer Mucicarmine Stain:

For storage requirements and expiration date refer to individual product labels.

APPLICATION:

Newcomer Supply Mucicarmine Stock Stain, Mayer a component of the Mucin, Mayer Mucicarmine Stain procedure, is used to stain acid epithelial mucin (sialomucin, sulfomucin) and also useful for the demonstration of the encapsulated yeast Cryptococcus neoformans. Metanil Yellow or Tartrazine Stain provides a light yellow counterstain to connective tissue and cytoplasmic elements.

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)
Technique:  Paraffin sections cut at 4 microns
Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

STAINING PROCEDURE:

1. 1. If necessary, heat dry tissue sections/slides in oven.
   2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
      1. 1. See Procedure Notes #1 and #2.
   3. Prepare fresh Weigert Iron Hematoxylin (Part 1409) Working Solution directly before use; combine and mix well.
      1. 1. Solution A: Ferric Chloride, Acidified   20 ml
         2. Solution B: Hematoxylin 1%, Alcoholic             20 ml
   4. Stain in Weigert Iron Hematoxylin Working Solution; 7 minutes.
   5. Rinse in running tap water for 10 minutes.
   6. Prepare Mayer Mucicarmine Working Solution; combine, mix well.
      1. 1. Mucicarmine Stock Stain, Mayer                 10 ml  
         2. Tap Water (do not use distilled water) 30 ml      
   7. Stain in Mayer Mucicarmine Working Solution for 60 minutes or longer if a more intense stain is desired.

Microwave Modification:  See Procedure Note #3.

1. 1. 1. 1. Place slides in a plastic Coplin jar containing Mayer Mucicarmine Working Solution. Microwave 10 minutes at 70°C.

1. 8. Rinse in several changes of tap water.
   9. Counterstain with preferred method:

• • 1. 1. Metanil Yellow Stain, Aqueous; stain 30 to 60 seconds. Proceed directly to Step #10.
       2. Tartrazine Stain 0.25%, Acetic Aqueous; 5 quick dips, rinse 30 seconds in distilled water. Proceed to Step #10.

1. 10. Dehydrate quickly through 95% and 100% ethyl alcohols. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

RESULTS:

PROCEDURE NOTES:

1. 1. Drain slides after each step to prevent solution carry over.
   2. Do not allow sections to dry out at any point during procedure.
   3. The microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

REFERENCES:

1. 1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 174-175.
   2. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 142-144.
   3. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 161-162.
   4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 168-169.
   5. Modifications developed by Newcomer Supply Laboratory.