Validation Stain: Gomori One-Step, Aniline Blue
Other Applicable Stains: Masson Trichrome and McLetchie Trichrome
Tissue: Positive staining uterus.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain: Gomori One-Step Aniline Blue 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 Trichrome, Gomori One-Step, Aniline Blue Stain Kit:||Part 9176B/A||Individual Stain Solution|
|Solution A:||Bouin Fluid||250/500 ml||Part 1020|
|Solution B:||Ferric Chloride, Acidified||125/250 ml||Part 1409|
|Solution C:||Hematoxylin 1%, Alcoholic||125/250 ml||Part 1409|
|Solution D:||Trichrome Stain, Gomori One-Step, Aniline Blue||250/500 ml||Part 1403|
|Solution E:||Acetic Acid 0.5%, Aqueous||250/500 ml||Part 100121|
Newcomer Supply Trichrome, Uterus Control Slides are for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.
- Heat dry sections in oven according to your laboratory protocol.
- Preheat Solution A: Bouin Fluid to 56-60°C in oven or water bath. (Skip if using overnight method or microwave procedure.)
NEWCOMER SUPPLY VALIDATION PROCEDURE:
- 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.
- Mordant in preheated Solution A: Bouin Fluid (Step #2) for one hour at 56-60°C or overnight at room temperature. Cool at room temperature for 5-10 minutes.
- Skip Step #4 if tissue was originally Bouin fixed.
Microwave Modification: See Procedure Note #3.
- Place slides in a plastic Coplin jar containing Solution A: Bouin Fluid and microwave for 5 minutes at 60°C.
- Wash well in running tap water; rinse in distilled water.
- Prepare fresh Weigert Iron Hematoxylin; combine and mix well.
- Solution B: Ferric Chloride, Acidified 20 ml
- Solution C: Hematoxylin 1%, Alcoholic 20 ml
- Stain in fresh Weigert Iron Hematoxylin for 10 minutes.
- Wash in running tap water for 10 minutes; rinse in distilled water.
- See Procedure Note #4.
- Stain with Solution D: Trichrome Stain, Gomori One-Step, Aniline Blue for 20 minutes.
- Differentiate in Solution E: Acetic Acid 0.5%, Aqueous; 2 minutes.
- Rinse quickly in distilled water.
- 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.
|Collagen and mucin||Blue|
|Muscle fibers, cytoplasm and keratin||Red|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- The suggested microwave procedure has been tested at Newcomer Supply. This procedure is a guideline and techniques should be developed for use in your laboratory.
- If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
- Trichrome, Uterus Control Slides are validated with Trichrome Stain Kit, Gomori One-Step, Aniline Blue but can be used as positive controls with any preferred Trichrome procedure.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Brown, Richard. Histologic Preparations: Common Problems and Their Solutions. Northfield, Ill.: College of American Pathologists, 2009. 95-101.
- Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 165-166.
- Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 191-192.
- Vacca, Linda L. Laboratory Manual of Histochemistry. New York: Raven Press, 1985. 308-310.
- Modifications developed by Newcomer Supply Laboratory.