Elastic, Skin

Controls Slides Histopathology Elastic VVG

Validation Stain: Verhoeff
Other Applicable Stains: Gomori Aldehyde Fuchsin and Orcein

 

Tissue: 
(+) Skin
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4195A10/Set $49.00
Part # 4195B98/Set $346.00

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining skin.

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

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Verhoeff-Van Gieson Elastic quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 

Storage: 15-30°C in a light deprived and humidity controlled environment.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

PRODUCT DESCRIPTION:

The enclosed positive control slides are intended to verify histological techniques and reagent reactivity. The intended use is for the qualitative purpose of determining positive or negative results, and not intended for any quantitative purpose. These positive control slides are produced from human surgical or autopsy tissues under carefully controlled conditions.  Quality control measures are used to deliver control slides that are as consistent as possible. 

 

CONTROL SLIDE VALIDATION:

With Elastic, Verhoeff Stain Kit:   Part 9116A/B Individual Stain Solution
Solution A: Hematoxylin 5%, Alcoholic 125/250 ml Part 11623
Solution B: Ferric Chloride 10%, Aqueous 125/250 ml Part 10856
Solution C: Iodine, Weigert & Lugol, Aqueous 75/150 ml Part 12092
Solution D: Sodium Thiosulfate 5%, Aqueous 250/500 ml Part 1389
Solution E: Van Gieson Stain 250/500 ml Part 1404

 

APPLICATION:

Newcomer Supply Elastic, Skin Control Slides are for the positive histochemical staining of elastic fibers in skin.

 

PRESTAINING PREPARATION:

  1. Heat dry sections in oven according to your laboratory protocol.
  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.
  6. Wash well in tap water.
  7. Place in Solution D: Sodium Thiosulfate 5%, Aqueous for 1 minute.
  8. Wash well in running tap water for 5 minutes.
  9. Counterstain in Solution E: Van Gieson Stain for 3 to 5 minutes.
  10. 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/nuclei 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.