Victoria Blue Stain, Alcoholic

(uses include: Hepatitis B surface antigen, copper and venous invasion.)

Product Options:

Part # 1406A 500 ml $167.30 H
Part # 1406C 1 L $304.00 H

SOLUTION:                                                                                                                    

500 ml 1 Liter
Victoria Blue Stain, Alcoholic Part 1406A Part 1406C

Additionally Needed:

Elastic, Aorta Control Slides

                 OR

Elastic, Skin Control Slides

Part 4194

      OR

Part 4195

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Potassium Permanganate 1%, Aqueous Part 13393
Sulfuric Acid 1%, Aqueous Part 14012
Sodium Bisulfite 1%, Aqueous Part 13821
Alcohol, Ethyl Denatured, 70% Part 10844
Nuclear Fast Red Stain, Kernechtrot Part 1255

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

 

APPLICATION:

Newcomer Supply Victoria Blue Stain, Alcoholic, a uniquely blended stain solution, is used for demonstration of connective tissue, elastic fibers and fibrosis.  Other applications include staining of copper-associated protein in liver sections.

 

METHOD:

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

Technique:  Paraffin sections cut at 5 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 staining procedure provided below.

 

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 Note #1.
  2. Prepare fresh Potassium Permanganate-Sulfuric Acid Working Solution; combine and mix well.
    1. Potassium Permanganate 1%, Aqueous (Part 13393)      10 ml
    2. Sulfuric Acid 1%, Aqueous (Part 14012)                          10 ml  
    3. Distilled Water                                                           40 ml  
  3. Place slides in Potassium Permanganate-Sulfuric Acid Working Solution for 5 minutes.
  4. Treat with Sodium Bisulfite 1%, Aqueous (Part 13821) for 2 minutes or until sections are colorless.
  5. Wash slides well in running tap water.
  6. Rinse in 70% ethyl alcohol (Part 10844) for 2 minutes.
  7. Stain in Victoria Blue Stain, Alcoholic for a minimum of 4 hours.
    1. See Procedure Note #2.
  8. Differentiate in 70% ethyl alcohol for 1-3 minutes or until background is completely decolorized.
  9. Wash slides well in running tap water.
  10. Counterstain in Nuclear Fast Red Stain, Kernechtrot (Part 1255) for 5 minutes.
  11. Wash in running tap water for 5 minutes.
  12. 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:

Elastic fibers Blue
Copper-associated protein Blue (if present in liver sections)
Nuclei and cytoplasm Red

 

PROCEDURE NOTES:

  1. Drain staining rack/slides after each step to prevent solution carry over.
  2. For best results, overnight staining at room temperature in Victoria Blue Stain, Alcoholic is recommended.
  3. 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. 296-297.
  2. Prophet, Edna B., Bob Mills, Jacquelyn Arrington, and Leslie Sobin. Laboratory Methods in Histotechnology. Washington, D.C.; American Registry of Pathology. 11992. 210-211.
  3. Tanaka, Kaoru, Wataru Mori, and Koji Suwa. “Victoria Blue-Nuclear Fast Red Stain for HBs Antigen Detection in Paraffin Section.”  Pathology International 31.1 (1981): 93-98.
  4. Tsutsumi, Yutaka, Noboru Onoda, and Yoshiyuki Osamura. “Victoria Blue-Hematoxylin and Eosin Staining: A Useful Routine Stain for Demonstration of Venous Invasion by Cancer Cells.” The Journal of Histotechnology 13.4 (1990): 271-274.
  5. Modifications developed by Newcomer Supply Laboratory.