Acid Fast Bacteria (AFB), Animal
Validation Stain: AFB, Ziehl-Neelsen
Other Applicable Stains: AFB, Kinyoun and Auramine/Rhodanine
Tissue: Positive staining animal organ 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: AFB, Ziehl-Neelsen 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 AFB, Ziehl-Neelsen Stain Kit:||Part 9101A||Individual Stain Solution|
|Solution A:||Carbol Fuchsin Stain, Ziehl-Neelsen||250 ml||Part 1030|
|Solution B:||Acid Alcohol 1%||250 ml||Part 10011|
|Solution C:||Light Green SF Yellowish Stain 0.1%, Aqueous||250 ml||Part 12203|
Newcomer Supply Acid Fast Bacteria (AFB), Animal Control Slides are for the positive histochemical staining of acid-fast mycobacteria in tissue sections.
- Heat dry sections in oven according to your laboratory protocol.
- Filter Solution A: Carbol Fuchsin Stain, Ziehl-Neelsen with filter paper whenever a thick sheen develops on solution surface.
NEWCOMER SUPPLY VALIDATION PROCEDURE:
- 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.
- See Procedure Notes #1 and #2.
- Stain in Solution A: Carbol Fuchsin Stain, Ziehl-Neelsen for 15 minutes at room temperature. Keep solution covered.
- See Procedure Note #3.
- Rinse in running tap water for 2 to 3 minutes.
- Differentiate in Solution B: Acid Alcohol 1% until color no longer runs off the slide and sections are pale pink; 3 to 10 rapid dips.
- Wash in running tap water 3 to 5 minutes; rinse in distilled water.
- Counterstain in Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous; 2-5 dips.
- Rinse with one quick dip in distilled water or proceed directly to Step #10 without a distilled water rinse.
- 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.
|Acid-fast bacilli||Bright red|
|Lung||Negative for AFB|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- Sections can remain in Carbol Fuchsin Stain, Ziehl-Neelsen for up to 60 minutes without adverse effect. Additional differentiation may be required in Step #6.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 218-220.
- Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 237.
- Modifications developed by Newcomer Supply Laboratory.