Trichrome, Multi-Tissue

Controls Slides Histopathology TRICHROME MULTI-TISSUE

Validation Stain: Gomori One-Step, Aniline Blue
Other Applicable Stains: Masson Trichrome and McLetchie Trichrome

 

Tissue: 
(+) Liver, (+) Kidney, (+) Uterus
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4693A10/Set $75.00
Part # 4693B98/Set $525.00

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver, positive staining kidney and 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.

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 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

 

APPLICATION:

Newcomer Supply Trichrome, Multi-Tissue Control Slides, use a combination of tissue sources for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.

 

PRESTAINING PREPARATION:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. 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:

  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. 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.
    1. Skip Step #4 if tissue was originally Bouin fixed.

       Microwave Modification:  See Procedure Note #3.

  1. Place slides in a plastic Coplin jar containing Solution A: Bouin Fluid and microwave for 5 minutes at 60°C.
  1. Wash well in running tap water; rinse in distilled water.
  2. Prepare fresh Weigert Iron Hematoxylin; combine and mix well.
    1. Solution B: Ferric Chloride, Acidified               20 ml
    2. Solution C: Hematoxylin 1%, Alcoholic            20 ml
  3. Stain in fresh Weigert Iron Hematoxylin for 10 minutes.
  4. Wash in running tap water for 10 minutes; rinse in distilled water.
    1. See Procedure Note #4.
  5. Stain with Solution D: Trichrome Stain, Gomori One-Step, Aniline Blue for 20 minutes.
  6. Differentiate in Solution E: Acetic Acid 0.5%, Aqueous; 2 minutes.
  7. Rinse quickly in distilled water.
  8. 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:

Collagen and mucin Blue
Muscle fibers, cytoplasm and keratin Red
Nuclei Blue/black

 

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. 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.
  4. If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
  5. Trichrome, Multi-Tissue Control Slides are validated with Trichrome Stain Kit, Gomori One-Step, Aniline Blue but can be used as positive controls with the end users Trichrome stain procedure of preference.
  6. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Brown, Richard. Histologic Preparations: Common Problems and Their Solutions. Northfield, Ill.: College of American Pathologists, 2009. 95-101.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 165-166.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 191-192.
  4. Vacca, Linda L. Laboratory Manual of Histochemistry. New York: Raven Press, 1985. 308-310.
  5. Modifications developed by Newcomer Supply Laboratory.