Colloidal Iron, Müller-Mowry Stain Kit

colloidal iron stain kit muller mowry

This procedure is used to demonstrate acid epithelial mucin (sialomucin, sulfomucin) and stromal (mesenchymal) mucin. This method is also excellent for the demonstration of the encapsulated yeast Cryptococcus neoformans.

Product Options:

Part # 9110A250ml kit $274.70H

 

  

COLLOIDAL IRON STAIN KIT INCLUDES:                      

    Part 9110A
Solution A: Acetic Acid 12%, Aqueous 500 ml x 2
Solution B: Colloidal Iron Stock 125 ml
Solution C: Acetic Acid, Glacial, ACS 50 ml
Solution D: Potassium Ferrocyanide 2%, Aqueous 125 ml
Solution E: Hydrochloric Acid 2%, Aqueous 125 ml
Solution F: Van Gieson Stain 250 ml

 

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed with this kit are two complimentary unstained positive control slides to be used for the initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and additional control slides may be available for purchase under separate part numbers.

Additionally Needed:

Hydrochloric Acid 5%, Aqueous Part 12086 (for acid cleaning glassware)
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

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

 

APPLICATION:

Newcomer Supply Colloidal Iron, Mϋller-Mowry Stain Kit procedure is used to demonstrate acid epithelial mucin (sialomucin, sulfomucin) and stromal (mesenchymal) mucin. This method is also excellent for the demonstration of the encapsulated yeast Cryptococcus neoformans.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

  • Chromate fixatives should be avoided

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

PRESTAINING PREPARATION:

  1. To avoid the possibility of residual background iron staining, acid clean glassware is recommended in the staining procedure.
    1. See Procedure Note #1.
  2. Prepare Colloidal Iron Working Solution; combine and mix well.
    1.   Solution B: Colloidal Iron Stock                          20 ml
    2.   Solution C: Acetic Acid, Glacial ACS                   5 ml
    3.   Distilled Water                                                    15 ml

 

STAINING PROCEDURE:

  1. 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. See Procedure Notes #2 and #3.
  2. Place in Solution A: Acetic Acid 12%, Aqueous for 30 seconds.
  3. Drain Slides. Do not rinse.
  4. Place in Colloidal Iron Working Solution (Step #2) for 30 minutes.
  5. Rinse in three changes of Solution A: Acetic Acid 12%, Aqueous; 3 minutes each.
  6. Prepare fresh Ferrocyanide-Hydrochloric Acid Solution directly before use; combine and mix well.
    1. Solution D: Potassium Ferrocyanide 2%, Aqueous  20 ml
    2. Solution E: Hydrochloric Acid 2%, Aqueous          20 ml
  1. Place in fresh Ferrocyanide-Hydrochloric Acid Solution for 15 minutes.
  2. Wash in running tap water for 1-5 minutes.
  3. Counterstain in Solution F: Van Gieson Stain for 3-5 minutes.
    1. Proceed directly to dehydration step without rinsing.
  4. Dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucin Blue
Stromal mucin Blue
Capsule of Cryptococcus neoformans Blue
Collagen Red
Muscle and cytoplasm Yellow

 

PROCEDURE NOTES:

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water. Cleaning glassware with bleach is not equivalent to acid washing.
  2. Drain staining rack/slides after each step to prevent solution carry over.
  3. Do not allow sections to dry out at any point during staining procedure.
  4. Nuclear Fast Red, Kernechtrot (Part 1255) can be used as an alternative counterstain.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 175-176.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 151-153.
  3. Rekhtman, Natasha, and Justin Bishop. Quick Reference Handbook for Surgical Pathologists. Berlin: Springer, 2011. 69.
  4. Modifications developed by Newcomer Supply Laboratory.