Phosphotungstic Acid Hematoxylin (PTAH)

Controls Slides Histopathology PHOSPHOTUNGSTIC ACID HEMATOXYLIN (PTAH)

Validation Stain: Phosphotungstic Acid Hematoxylin (PTAH)

 

Tissue: 
(+) Striated Muscle
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4565A10/Set $49.00
Part # 4565B98/Set $346.00

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining striated muscle.

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

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

Quality Control Stain:  PTAH 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 Phosphotungstic Acid Hematoxylin (PTAH) Stain Kit:  Part 9111A Individual Stain Solution
Solution A: Zenker Fixative, Modified, Zinc Chloride 250 ml Part 1461
Solution B: Acetic Acid, Glacial, ACS 25 ml Part 10010
Solution C: Potassium Permanganate 0.25%, Aqueous 250 ml Part 133931
Solution D: Oxalic Acid 5%, Aqueous 250 ml Part 1293
Solution E: Phosphotungstic Acid Hematoxylin (PTAH) Stain, Modified Mallory 250 ml Part 1334

 

APPLICATION:

The Newcomer Supply Phosphotungstic Acid Hematoxylin (PTAH) Control Slides are for the positive histochemical demonstration of muscle striations and collagen in tissue sections.

 

PRESTAINING PREPARATION:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Prepare Zenker Fixative Working Solution; combine and mix well.

        Solution A: Zenker Fixative, Modified, Zinc Chloride       38 ml

        Solution B: Acetic Acid, Glacial, ACS                                2 ml

 

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. Fix slides in Zenker Fixative Working Solution (Step #2) at 56°C for 3 hours.

        Microwave Modification:  See Procedure Note #3.

  1. Place slides in a plastic Coplin jar containing prepared Zenker Fixative Working Solution and microwave for 5 minutes at 60°C.
  1. Wash well in three changes of tap water; rinse in distilled water.
  2. Place in Solution C: Potassium Permanganate 0.25%, Aqueous for 10 minutes.
  3. Wash in three changes of tap water; rinse in distilled water.
  4. Place in Solution D: Oxalic Acid 5%, Aqueous for 10 minutes.
  5. Wash in three changes of tap water; rinse in distilled water.
  6. Stain in Solution E: PTAH Stain, Modified Mallory for 12-24 hours at room temperature, or 2 hours at 56°C.
    1. See Procedure Note #4.

        Microwave Modification:

  1. Place slides in a plastic Coplin jar containing Solution E: PTAH Stain, Modified Mallory and microwave for 7 minutes at 70°C.
  1. Dehydrate quickly in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
    1. Dehydrate quickly as alcohol may extract stain from sections.

 

RESULTS:

Muscle striations, fibrin, keratin Dark blue
Collagen, cartilage, elastic fibers Deep reddish brown
Nuclei Blue

 

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. Newcomer Supply PTAH Stain formula is twice as strong as the original Mallory formulation; adjust staining time according to preference of intensity.  Suggested staining time at 37°C is 18 hours.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008.130-131.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 178-180, 201-202.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 193-194.
  4. Modifications developed by Newcomer Supply Laboratory.