Elastic, Verhoeff Stain Kit

elastic stain kit verhoeff

Product Options:

Part # 9116A250ml kit $192.60H
Part # 9116B500ml kit $259.50H

 

  

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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 Solution 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 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:

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 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 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. 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.
  1. Prepare fresh Verhoeff Working Solution by combining in the order listed, mixing well after each addition. This is essential for optimal staining.
    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
  1. Stain in fresh Verhoeff Working Solution for 1 hour.
  1. Discard solution after successful differentiation in Step 6.
  1. Rinse in several changes of tap water.
  2. Prepare fresh Ferric Chloride 2%, Aqueous Solution:
  1. Solution B: Ferric Chloride 10%, Aqueous      10 ml
  2. Distilled Water                                                        40 ml
  1. Differentiate each slide individually in fresh Ferric Chloride 2%, Aqueous Solution with agitation; approximately 30 dips.
  2. Check differentiation; rinse well in tap water and check microscopically for black elastic staining with gray background.  Repeat in Ferric Chloride, 2% Aqueous Solution if necessary until desired elastic differentiation is achieved.  Repeat steps for each slide.
  1. See Procedure Notes #3 and #4.
  1. Wash well in tap water.
  2. Place slides 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.
  1. See Procedure Note #5.
  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:

Elastic fibers/tissue Blue-black to black
Nuclei Blue to black
Collagen Red
Other tissue elements Yellow

                              

PROCEDURE NOTES:

  1. Drain staining rack/slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during staining procedure.
  3. It is easy to over-differentiate in Ferric Chloride 2%, Aqueous. If the background is completely colorless, the section has been over-differentiated. Over-differentiated sections may be re-stained in Verhoeff Working Solution (Step #3) provided sections have not been treated with an alcohol step.
  4. For optimal results, slides must be individually differentiated. Timing of each slides differentiation can vary dependent upon the amount of elastic tissue present in sections.
  5. Do not prolong staining in Solution E: Van Gieson Stain. The picric acid element will act to further differentiate the stain.
  6. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida, Histotechnology: A Self-Instructional Text. 2nd ed. Chicago: ASCP Press, 1997. 138-140.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 196-197.
  3. Modifications developed by Newcomer Supply Laboratory.