Helicobacter, Toluidine Blue / Alcian Yellow Stain Kit

Used to screen and detect Helicobacter pylori in gastrointestinal biopsy specimens.

Product Options:

Part # 9130A 250ml kit $253.80 H

 

  

HELICOBACTER, TOLUIDINE BLUE/ALCIAN YELLOW STAIN KIT INCLUDES:

Part 9130A 
Solution A: Periodic Acid 1%, Aqueous 250 ml
Solution B: Sodium Metabisulfite 5%, Acidified 250 ml
Solution C: Alcian Yellow Stain 1%, Alcoholic 250 ml
Solution D: Toluidine Blue Stock Stain 1%, Aqueous 10 ml
Solution E: Sodium Hydroxide 3%, Aqueous 10 ml

 

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed are two complimentary unstained positive control slides 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 control slides may be available for purchase under separate part numbers.

 

Additionally Needed:

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 Helicobacter, Toluidine Blue/Alcian Yellow Stain Kit procedure is used for screening and detection of Helicobacter pylori in gastrointestinal biopsy specimens.

 

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 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.

 

STAINING PROCEDURE:                  

    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. See Procedure Note #1.
    3. Oxidize in Solution A: Periodic Acid 1%, Aqueous; 10 minutes.
    4. Wash slides well with tap water.
    5. Place in Solution B: Sodium Metabisulfite 5%, Acidified; 5 minutes.
    6. Wash in tap water for 5 minutes.
    7. Stain in Solution C: Alcian Yellow Stain 1%, Alcoholic; 5 minutes.
    8. Wash well in tap water.
    9. Prepare fresh Toluidine Blue Working Solution; combine, mix well.
        1. Distilled Water                                                              50 ml
        2. Solution D: Toluidine Blue Stock Stain 1%, Aqueous 5 ml
        3. Solution E: Sodium Hydroxide 3%, Aqueous        2 drops
    10. Stain in fresh Toluidine Blue Working Solution; 3 minutes.
    11. Wash well in tap water.
    12. Allow slides to air-dry completely.
    13. Clear dried slides in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
        1. Eliminate alcohol dehydration to retain organism staining.

 

RESULTS:

Helicobacter pylori Blue
Mucin Yellow
Background Pale blue

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 225-226.
    2. Leung, Jennifer, Kevin Gibbon and Robert Vartaniam. “Rapid Staining Method for Helicobacter pylori in Gastric Biopsies.” The Journal of Histotechnology 12 (1996): 131-132.
    3. Modifications developed by Newcomer Supply Laboratory.