Carbol Fuchsin Stain, Kinyoun

(use: Kinyoun for AFB)

Product Options:

Part # 1031A 250 ml $50.80 H
Part # 1031B 500 ml $94.40 H

SOLUTION:                                                                                                       

250 ml 500 ml
Carbol Fuchsin Stain, Kinyoun Part 1031A Part 1031B

 

Additionally Needed:

Acid Fast Bacteria (AFB) Control Slides Part 4011
Acid Alcohol 1% Part 10011
Methylene Blue Stain 0.14%, Alcoholic

OR

Methylene Blue Stain 1.4%, Alcoholic

Part 12401A

    OR

Part 1240A

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Carbol Fuchsin Stain, Kinyoun a crucial element in the AFB Stain, Kinyoun procedure is used to demonstrate the presence of acid-fast mycobacteria in tissue sections. Phenol is employed in this solution to render the cell walls of bacteria permeable to the fuchsin stain.  The use of weak acid for differentiation allows excess stain to be removed from tissues, but will not remove stain from the acid-fast organisms.

 

METHOD:

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

Technique:  Paraffin sections cut at 4 microns

Solutions: All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Filter Carbol Fuchsin Stain, Kinyoun with filter paper before use.

 

STAINING 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. Stain in freshly filtered Carbol Fuchsin Stain, Kinyoun for 60 minutes at room temperature. Keep solution covered.
  3. Wash in running tap water for 2 to 3 minutes.
  4. Differentiate in Acid Alcohol 1% (Part 10011) until color no longer runs off the slide and sections are pale pink; 3 to 10 rapid dips.
  5. Wash in running tap water 3 to 5 minutes; rinse in distilled water.
  6. Counterstain in Methylene Blue Stain 0.14%, Alcoholic (Part 12401). Or dilute 5 ml Methylene Blue Stain 1.4%, Alcoholic (Part 1240) with 45 ml tap water to a 0.14% solution.
    1. Counterstain with 3-6 dips.
    2. Rinse in tap water, followed by a distilled water rinse.
    3. Sections should be pale blue.
    4. See Procedure Notes #3 and #4.
  7. Rinse in distilled water.
  8. 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. See Procedure Note #5.

 

RESULTS:

Acid-fast bacteria Bright red
Background Pale 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. If section is over-stained, remove methylene blue with Acid Alcohol 1% (Part 10011), rinse thoroughly, and repeat methylene blue counterstain step (Step #8) with fewer dips.
  4. Light Green SF Yellowish Stain 0.1%, Aqueous (Part 12203) can be used as a counterstain in place of Methylene Blue.
    1. Stain for 3-6 dips.
    2. Rinse with one quick dip in distilled water or proceed directly to Step #10 without a distilled water rinse.  
    3. Background will be green.
  5. Dehydrate quickly to maintain methylene blue staining.
  6. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 224-226.
  2. Kinyoun. J.J. “A Note on Uhlenhuths Method for Sputum Examination, for Tubercle Bacilli.” American Journal of Public Health 5.9 (1915). 867-870.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 236-237.
  4. Modifications developed by Newcomer Supply Laboratory.