AFB STAIN KIT, Fite

Description

AFB STAIN KIT, Fite

AFB STAIN KIT INCLUDES:                         

AFB Stain Kit bottles are not available to be purchased individually.  "Component" bottles may be available under separate part numbers.

For storage temperature refer to individual bottle labels.

AFB STAIN KIT APPLICATION:

To detect the presence of Nocardia or Mycobacterium leprae (causative agent of leprosy) in tissue sections with minor variations in the procedure.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

AFB STAIN KIT METHOD:

Fixation:          10% Neutral Buffered Formalin

Technique:      Paraffin sections cut at 5 microns

Solutions:        All solutions manufactured by Newcomer Supply

AFB STAIN KIT PROCEDURE:

1. Filter Solution B, Carbol Fuchsin Stain Solution, Ziehl-Neelsen.
2. Deparaffinize slides in Solution A, Xylene/Peanut Oil, two changes, 12 minutes each.
3. Drain slides, wipe off excess oil, and blot to opacity taking care to remove residual oil. 
4. Stain slides in freshly filtered Solution B, Carbol Fuchsin Stain Solution for 30 minutes at room temperature.
5. Wash in running tap water for 3 minutes.
6. If staining for Nocardia, differentiate slides in Solution C, Sulfuric Acid 1% Aqueous Solution for 3 minutes. (Mycobacterium leprae slides should be differentiated individually until sections are light pink. This should take about 1 minute.)
7. Wash in running tap water for 3 minutes.
8. Counterstain lightly with Solution D, Methylene Blue Stain Solution, 0.5% for 5-10 seconds. (See procedure notes.)
9. Rinse off excess Methylene Blue Stain Solution, 0.5% in running tap water.  Background should be a light sky blue.
10. Blot and let air-dry completely.
11. Dip slides in xylene and coverslip with a compatible mounting medium.

AFB STAIN KIT RESULTS:

AFB STAIN KIT PROCEDURE NOTES:

1. A small percentage of Nocardia organisms may resist taking the red stain and remain blue due to the growth phase of the individual organism.
2. Acid fastness of the leprosy organisms is enhanced when the waxy capsule is protected by the mixture of xylene-peanut oil and the avoidance of dehydrating solutions.
3. It is very important to remove excess peanut oil from tissue, or it will have a blotchy appearance.  To remove as much oil as possible, place slide face down on clean paper towel.  While holding in place, wipe gently but firmly the back of the slide. Transfer slide to another paper towel and repeat this step to ensure oil is removed properly.
4. Do not over-stain with methylene blue, as the organisms may be masked. Check microscopically before air drying. If overstained, remove methylene blue with a dilute acid solution, rinse thoroughly, then repeat step 7 with a shorter timing.   
5. The methylene blue may stain paler than usual if the tap water used has an acidic pH.

COMPLIMENTARY UNSTAINED POSITIVE CONTROL SLIDES:

Each kit comes with two complimentary unstained positive control slides to be used for the initial verification of staining technique and reagents.  We suggest running one slide along with your in-house positive control slide for the first run.  Retain the second slide for future trouble shooting, if needed.

REFERENCES:

1. "Histopathological Methods and Color Atlas of Special Stains and Tissue Artifacts", 1992, Luna, Lee, Johnson Printers, Downers Grove, Illinois, 1992, pp. 180-181.
2. "Histotechnology: a Self-Instructional Text", Carson, 2009, 3^rd Ed., pp. 228-229.
3. "Theory and Practice of Histotechnology", 1980, Sheehan, D., and Hrapchak, B., 2^nd edition, Batelle Press, Columbus and Richland, p. 237.