Iron, Gomori Prussian Blue Stain Kit

iron stain kit gomori prussian blue

Product Options:

Part # 9136A250ml kit $123.60H
Part # 9136B500ml kit $190.00H

 

  

IRON STAIN KIT INCLUDES:

    Part 9136A Part 9136B
Solution A: Hydrochloric Acid 20%, Aqueous 125 ml 250 ml
Solution B: Potassium Ferrocyanide 10%,  Aqueous 125 ml 250 ml
Solution C: Nuclear Fast Red Stain, Kernechtrot 250 ml 500 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 Iron, Gomori Prussian Blue Stain Kit procedure is used to detect loosely bound ferric iron in tissue sections. This histochemical reaction is sensitive enough to demonstrate even minute amounts of iron deposits in blood cells, bone marrow and spleen. 

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090) (Chromate fixatives should be avoided)

Technique:   Paraffin sections cut at 5 microns

Solutions:     All solutions 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.

 

PROCEDURE:

  1. To avoid the possibility of residual background iron staining, acid clean glassware is recommended in the staining procedure.
  1. See Procedure Note #1.
  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.
  1. Prepare fresh Ferrocyanide Working Solution directly before use; combine and mix well:
  1. Solution A: Hydrochloric Acid 20%, Aqueous    20 ml
  2. Solution B: Potassium Ferrocyanide 10%, Aqueous         20 ml
  1. Place slides in fresh Ferrocyanide Working Solution for 20 minutes.
  2. Rinse in three changes of tap water; rinse in distilled water.
  3. Place slides in Solution C: Nuclear Fast Red Stain, Kernechtrot  for 5 minutes.
  1. Shake solution well before use; do not filter.
  1. Rinse well in distilled water.
  1. See Procedure Note #4.
  1. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.  

 

RESULTS:

Ferric Iron Deposits Bright Blue
Nuclei Red
Cytoplasm Pink

 

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. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 179-184.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 217-218.
  3. Modifications developed by Newcomer Supply Laboratory.