Validation Stain: Verhoeff
Other Applicable Stains: Gomori Aldehyde Fuchsin and Orcein
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.
Intended Use: To verify histological techniques and reagent reactivity.
Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.
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|
Newcomer Supply Elastic, Skin Control Slides are for the positive histochemical staining of elastic fibers in skin.
- Heat dry sections in oven according to your laboratory protocol.
- Prepare fresh Verhoeff Working Solution by combining in the exact order listed, mixing well after each addition. Save for Step #5.
- Solution A: Hematoxylin 5%, Alcoholic 20 ml
- Solution B: Ferric Chloride 10%, Aqueous 8 ml
- Solution C: Iodine, Weigert & Lugol, Aqueous 8 ml
- Prepare fresh Ferric Chloride 2%, Aqueous Solution for Step #7.
- Solution B: Ferric Chloride 10%, Aqueous 10 ml
- Distilled water 40 ml
- 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.
- See Procedure Notes #1 and #2.
- Stain in fresh Verhoeff Working Solution (Step #2) for 20 minutes.
- Discard solution after successful differentiation in Step #7.
- Rinse in several changes of tap water.
- Differentiate each slide individually in fresh Ferric Chloride 2%, Aqueous Solution (Step #3) with agitation; approximately 20 dips.
- 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.
- See Procedure Notes #3 and #4.
- Wash well in tap water.
- Place in Solution D: Sodium Thiosulfate 5%, Aqueous for 1 minute.
- Wash well in running tap water for 5 minutes.
- Counterstain in Solution E: Van Gieson Stain for 3 to 5 minutes.
- 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.
|Elastic fibers/tissue/nuclei||Blue-black to black|
|Other tissue elements||Yellow|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- It is easy to over-differentiate in Ferric Chloride 2%, Aqueous Solution.
- If background is completely colorless, the section has been over-differentiated.
- Over-differentiated sections may be re-stained in Step #5 provided sections have not been treated with alcohol.
- Slides must be individually differentiated. Differentiation can vary dependent upon the amount of elastic tissue present in sections.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 167-169.
- Mallory, Frank Burr, and James Homer Wright. Pathological Technique. 7th ed. Philadelphia, PA: W.B. Saunders Company, 1918. 118-119.
- Modifications developed by Newcomer Supply Laboratory.