Elastic, Verhoeff Stain Kit

Used to demonstrate pathologic changes in elastic fibers as well as demonstration of normal elastic tissue such as arteries and veins.

Product Options:

Part # 9116A 250ml kit $195.70 H
Part # 9116B 500ml kit $263.00 H

  

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ELASTIC, VERHOEFF STAIN KIT INCLUDES:

Part 9116A Part 9116B
Solution A: Hematoxylin 5%, Alcoholic 125 ml 250 ml
Solution B: Ferric Chloride 10%, Aqueous 125 ml 250 ml
Solution C: Iodine, Weigert & Lugol, Aqueous 75 ml 150 ml
Solution D: Sodium Thiosulfate 5%, Aqueous 250 ml 500 ml
Solution E: Van Gieson Stain 250 ml 500 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 additional 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 Elastic, Verhoeff Stain Kit procedure, commonly referred to as Verhoeff-Van Gieson (VVG) technique, is used to demonstrate pathologic changes in elastic fibers as well as demonstration of normal elastic tissue such as arteries and veins.

 

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.

 

PRESTAINING PREPARATION:

    1. If necessary, heat dry tissue sections/slides in oven.
    2. Prepare fresh Verhoeff Working Solution by combining in the exact order listed, mixing well after each addition. Save for Step #5.
        1. Solution A: Hematoxylin 5%, Alcoholic                       20 ml
        2. Solution B: Ferric Chloride 10%, Aqueous         8 ml
        3. Solution C: Iodine, Weigert & Lugol, Aqueous         8 ml
    3. Prepare fresh Ferric Chloride 2%, Aqueous Solution for Step #7.
        1. Solution B: Ferric Chloride 10%, Aqueous       10 ml
        2. Distilled water       40 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 #1 and #2.
    2. Stain in fresh Verhoeff Working Solution (Step #2) for 20 minutes.
        1. Discard solution after successful differentiation in Step #7.
    3. Rinse in several changes of tap water.
    4. Differentiate each slide individually in fresh Ferric Chloride 2%, Aqueous Solution (Step #3) with agitation; approximately 20 dips.
    5. Check differentiation: rinse well in tap water and check microscopically for black elastic staining with gray background.
        1. Repeat in Ferric Chloride 2%, Aqueous Solution if necessary until desired elastic differentiation is achieved.
        2. See Procedure Notes #3 and #4.
    1. Wash well in tap water.
    2. Place in Solution D: Sodium Thiosulfate 5%, Aqueous for 1 minute.
    3. Wash well in running tap water for 5 minutes.
    4. Counterstain in Solution E: Van Gieson Stain for 3 to 5 minutes.
    5. 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/tissue Blue-black to black
Collagen Red
Other tissue elements Yellow

 

PROCEDURE NOTES:

    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. It is easy to over-differentiate in Ferric Chloride 2%, Aqueous Solution.
        1. If background is completely colorless, the section has been over-differentiated.
        2. Over-differentiated sections may be re-stained in Step #5 provided sections have not been treated with alcohol.
    4. Slides must be individually differentiated. Differentiation can vary dependent upon the amount of elastic tissue present in sections.
    5. 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. 167-169.
    2. Mallory, Frank Burr, and James Homer Wright. Pathological Technique. 7th ed. Philadelphia, PA: W.B. Saunders Company, 1918. 118-119.
    3. Modifications developed by Newcomer Supply Laboratory.