Phosphotungstic Acid Hematoxylin (PTAH)

Phosphotungstic Acid Hematoxylin (PTAH) Stained Histology Slide

Validation Stain: Phosphotungstic Acid Hematoxylin (PTAH)

 

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

Product Options:

Part # 4565A 10/set $60.80
Part # 4565B 98/set $452.40

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.
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 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.
      1. Solution A: Zenker Fixative, Modified, Zinc Chloride       38 ml
      2. 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 in Zenker Fixative Working Solution (Step #2) at 56°C; 3 hours.

        Microwave Modification:  See Procedure Note #3.

        1. Place slides in a plastic Coplin jar containing Zenker Fixative Working Solution. Microwave 5 minutes at 60°
    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°
        1. See Procedure Note #4.

Microwave Modification:

        1. Place slides in a plastic Coplin jar containing Solution E: PTAH Stain, Modified Mallory. Microwave 7 minutes at 70°
    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, 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
    4. Adjust staining time according to preference of intensity. Suggested staining time at 37°C is 18 hours.
    5. If using a xylene substitute, follow manufacturer’s recommendation 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.