Fungus, PAS, Aspergillus sp., Artificial

Validation Stain: PAS/Light Green

 

(Hyphae forming fungus. Not in human tissue.)

Tissue: 
(+) Rat Lung / (-) Lung
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4232A 10/set $52.00
Part # 4232B 98/set $365.00

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat lung and negative staining human lung.

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

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

Quality Control Stain:  PAS/Light Green 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 Fungus, PAS/Light Green Stain Kit: Part 9122A Individual Stain Solution
Solution A: Periodic Acid 0.5%, Aqueous 250 ml Part 13308
Solution B: Schiff Reagent, McManus 250 ml Part 1371
Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous 250 ml Part 12203

   

APPLICATION:

Newcomer Supply Fungus, PAS, Aspergillus sp., Artificial Control Slides are for the positive histochemical staining of Aspergillus sp. in tissue sections. Aspergillus fumigatus purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. 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.
  3. Place in Solution A: Periodic Acid 0.5%, Aqueous for 5 minutes.
  4. Wash in three changes of tap water; rinse in distilled water.
  5. Drain slides of excess water and stain in Solution B: Schiff Reagent, McManus for 20 minutes.
  6. Wash gently in lukewarm tap water for 10 minutes to allow pink color to develop.
  7. Counterstain in Solution C:  Light Green SF Yellowish Stain 0.1%, Aqueous for 5 seconds.
    1. See Procedure Note #3.
  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:

Aspergillus sp.  Red to magenta
Background Pale green
Negative lung Negative for fungus

 

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. Increase or decrease staining time in Light Green SF Yellowish Stain 0.1%, Aqueous for preference of counterstain intensity.
  4. 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. 321-323.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245.
  3. Modifications developed by Newcomer Supply Laboratory.