Trichrome, Kidney

Controls Slides Histopathology Trichrome

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

 

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

Product Options:

Part # 4691A10/Set $46.00
Part # 4691B98/Set $330.00

Trichrome, Kidney Control Slides contain sections of positive staining kidney.

 

PRODUCT DESCRIPTION:

The enclosed positive control slides are intended to be used to verify histological techniques and reagent reactivity.  These slides are to be used for the qualitative purpose of determining positive or negative results, and are not intended to be used for any quantitative purpose.  The first serial section within the control box is stained and provided for your reference.  Before using the unstained slides, review the enclosed stained slide with your pathologist to ensure that this tissue source is acceptable.  Newcomer Supply will not accept a return with missing slides in the series. Newcomer Supply guarantees reactivity of these control slides for one year from the date of receipt.  Revalidate after one year to verify continued reactivity.  Store at 15-30°C in a light deprived and humidity controlled environment.

These positive control slides were produced from human surgical or autopsy tissues under carefully controlled conditions.  Reasonable measures are used to deliver quality control slides that are as consistent as possible.  However, characteristics of quality control slides may be dissimilar due to variations in the reagents, stains, techniques, laboratory conditions, and tissue sources used. Newcomer Supply Laboratory uses a manual method of performing quality control procedures, specifically avoiding automation, in order to provide reactive control slides for even less aggressive methods of staining that our customers may be using.

 

CONTROL SLIDE VALIDATION:                                        

With Trichrome, Gomori One-Step, Aniline BlueStain Kit: Part 9176A/B Individual Stain Solution
Solution A: Bouin Fluid 250/500 ml Part 1020
Solution B: Ferric Chloride, Aqueous 125/250 ml Part 1409 (Part of Set)
Solution C: Hematoxylin Stain 1%, Alcoholic 125/250 ml Part 1409 (Part of Set)
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

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

Newcomer Supply Trichrome, Kidney Control Slides are for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.

 

METHOD:

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

Technique:  Paraffin sections cut at 5 microns on Superfrost® Plus

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

 

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 Solution A: Bouin Fluid for one hour at 56-60°C or overnight at room temperature. Cool at room temperature for 5-10 minutes.
    1. Skip Step #2 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, Aqueous              20 ml
    2. Solution C: Hematoxylin 1%, Alcoholic            20 ml
  3. Stain slides 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.
  1. Differentiate in Solution E: Acetic Acid 0.5%, Aqueous for 2 minutes.
  2. Rinse quickly in distilled water.
  3. 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 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. The suggested microwave procedure has been tested at Newcomer Supply using an “EB Sciences”, 850 watt microwave oven with temperature probe and agitation tubes.  This procedure is reproducible in our laboratory.  It is nonetheless a guideline and techniques should be developed for your laboratory which meet the requirements of your situation. Microwave devices should be placed in a fume hood or vented into a fume hood, according to manufacturer’s instructions, to prevent exposure to chemical vapors.
  4. If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
  5. These Trichrome, Kidney Control Slides are validated with Trichrome Stain Kit, Gomori One-Step, Aniline Blue but can also 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.