ACID FAST STAIN: KINYOUN MODIFICATION

Technical Data #KINYOUNa / 98.07.22

The Kinyoun Modification Stain is used to stain acid-fast organisms as well as Mycobacterium sp. in specimens and in culture. Kinyoun modified Ziehl's hot procedure in such a manner that cold stain is used to wax content of Mycobacterium. Acid fast microorganisms are resistant to acid decolorizer, with a remaining reddish color stained and blue background.

FORMULA
in grams per liter

Carbol Fuchsine, Stain
Basic Fuchsine	40,0
Phenol	80,0
Ethanol, Denatured	200,0 ml
Water, deionized	1000,0 ml
Acid-Alcohol, Decolorizer
Ethanol, denatured	970,0 ml
Hydrochloric Acid, concentrated	30,0 ml
Methylene Blue, Counterstain
Methylene Blue	3,0
Water, deionized	1000,0 ml

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store in tightly-sealed containers at 15-30ºC in the dark.

SIGN OF DETERIORATION
Stains should not be used if they show signs of deterioration or if the expiration date has passed.

PROCEDURE

1. Prepare the smears as described in previous standardized methods directly from specimens or from an overnight growth.
2. Fix smears by passing through a Bunsen burner for 10 seconds.
3. Flood slides with Carbol Fuchsine and stain for 5 minutes.
4. Rinse thouroughly with tap water and decolorize with Acid-Alcohol until pink color flow disappears.
5. Rinse with tap water and add counterstain with Methylene Blue for 30 seconds.
6. Rinse with tap water. Blot dry and examine microscopically.

QUALITY CONTROL

Organisms	ATCC	Color	Morphology
Escherichia coli	25922	blue	bacilli
Mycobacterium tuberculosis	25177	reddish	large bacilli

LIMITATIONS OF METHOD
Slides of acid-fast should be stained individually to prevent acid fast bacilli carry-over from one slide to another. This is a part of identification. Other methods may be required.

REFERENCES

1. Finegold, S.M., E.J. Baron 1986 Bailey and Scott's Diagnostic Microbiology 7th ed. C.V. Mosby St. Louis
2. Lennette, E.H., A. Balows, W.J. Hausler Jr., H.J. Shadomy 1985 Manual of Clinical Microbiology 4th ed. American Society for Microbiology, Washington, D.C.
3. Vestal, A.L., 1969 Procedures for the isolation and identification of mycobacteria. USDHEW, Public Health Service No. 1995, U.S. Government Printing. Washington, D.C.

CATALOGUE NUMBERS

Basic Fuchsine	1 Kg	QB-46-0714
Methylene Blue	1 Kg	QB-46-3004
Solutions	500 ml	1 Liter	4 Liters	6x250 ml
Carbol fuchsine	8498	8548	8549	8499
Decolorizer	8485	8486	8487	8488
Methylene Blue	8475	8476	8477	8478
Complete Set	3 x 100 ml	8530
	3 x 250 ml	8550

2331, Dandurand	Phone	Fax	Internet
Montreal (Quebec)	(514) 277-2558	(514) 277-4714	http://www.quelab.com
Canada H2G 3C5	1 (800) 579-0998	1 (800) 579-8177	info@quelab.com