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FITES

The document outlines the procedure for Fite's Acid Fast Stain to demonstrate Mycobacterium leprae, detailing the necessary reagents, equipment, and safety precautions. It includes a step-by-step procedure for staining, with results indicating acid-fast bacilli appear red against a blue background. Additionally, it emphasizes the importance of using protective equipment due to the hazardous nature of some chemicals involved.

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6 views4 pages

FITES

The document outlines the procedure for Fite's Acid Fast Stain to demonstrate Mycobacterium leprae, detailing the necessary reagents, equipment, and safety precautions. It includes a step-by-step procedure for staining, with results indicating acid-fast bacilli appear red against a blue background. Additionally, it emphasizes the importance of using protective equipment due to the hazardous nature of some chemicals involved.

Uploaded by

drmillat140124
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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SURGICAL PATHOLOGY HISTOLOGY Date:

STAINING MANUAL - MICROORGANISMS Page: 1 of 2


FITE'S ACID FAST STAIN - LEPROSY
PURPOSE: To demonstrate mycobacterium leprae (leprosy), which are acid
fast organisms.

PRINCIPLE: This technique combines peanut oil with the deparaffinizing


solvent (xylene), minimizing the exposure of the bacteria's cell wall to
organic solvents, thus protecting the precarious acid-fastness of the
organism.

CONTROL: Leprosy positive tissue.

FIXATIVE: 10% formalin

TECHNIQUE: Cut paraffin sections 4-5 microns.

EQUIPMENT: Rinse all glassware in DI water, coplin jars, bibulous


blotting paper.

REAGENTS:
Xylene/Peanut Oil Solution: Ziehl-Neelsen Carbol-Fuchsin:
Xylene 50.0 ml See Ziehl-Neelsen AFB
Peanut Oil 50.0 ml
1% Acid Alcohol:
Mix well. Label with date and
See Ziehl-Neelsen AFB
initials, solution is stable for 1
year.
Working Methylene Blue:
CAUTION: Flammable, irritant. See Ziehl-Neelsen AFB

SAFETY/PPE: Wear nitrile gloves, goggles and lab coat. Keep hot,
uncapped, solutions in the exhaust hood. Avoid contact and inhalation.

Basic fuchsin: possible carcinogen.

Phenol: toxic by ingestion, inhalation and skin absorption. Readily absorbed


through skin, causing increased heart rate, convulsions and death. Will
burn eyes and skin, analgesic action may cause loss of pain. Target organ
effects in digestive, nervous and urinary systems.
MICROOGANISMS
FITE'S Page: 2 of 2
Hydrochloric acid: strong irritant to skin, eyes and respiratory system.

Target organ effects via inhalation on skin, respiratory, reproductive and


fetal systems.

Methylene blue: produced deleterious effects on fertility in rats.

PROCEDURE:
1. Deparaffinize in xylene/peanut 5. Wash in tap water.
oil mixture, 2 changes, 10 6. Acid alcohol until pale pink, dip
minutes each. until stain stops running.
2. Drain slides, blot off excess 7. Wash in tap water.
oil. 8. Counterstain in Working
3. Rinse in distilled water until Methylene blue, 30 seconds.
slide clears. 9. Wash in tap water.
4. Carbol-fuchsin, 30 minutes, 10. Blot and air dry.
room temperature. 11. Dip in xylene and coverslip.

RESULTS:
Acid-fast bacilli red
Background blue

NOTE: Mineral oil may be substituted for peanut oil.

REFERENCES:
Sheehan D, Hrapchak B, Theory and practice of Histechnology, 2nd Ed,
1980, pp237, Battelle Press, Ohio
Bancroft J, Stevens A, Theory and Practice of Histological Techniques, 2nd
Ed, pp 285, Churchill-Livingstone, NY
Crookham,J, Dapson,R, Hazardous Chemicals in the Histopathology
Laboratory, 2nd ED, 1991, Anatech

Prepared: By:

Approved: By:
Downloaded from WebPath: Internet Pathology Laboratory
http://www-medlib.med.utah.edu/WebPath/webpath.html
PROCEDURE CARD
FITE'S ACID FAST STAIN - LEPROSY
CONTROL: Leprosy positive tissue.

PROCEDURE:
1. Deparaffinize in xylene/peanut oil mixture, 2 changes, 10 minutes each.
2. Drain slides, blot off excess oil.
3. Rinse in distilled water until slide clears.
4. Carbol-fuchsin, 30 minutes, room temperature.
5. Wash in tap water.
6. Acid alcohol until pale pink, dip until stain stops running.
7. Wash in tap water.
8. Counterstain in Working Methylene blue, 30 seconds.
9. Wash in tap water.
10. Blot and air dry.
11. Dip in xylene and coverslip.

RESULTS:
Acid-fast bacilli red
Background blue

SAFETY: Keep hot solutions under fume hood.


Basic fuchsin: carcinogen

Xylene/Peanut Oil Solution: Ziehl-Neelsen Carbol-Fuchsin:


Xylene See Ziehl-Neelsen AFB
50.0 ml
P e a n u t 1% Acid Alcohol:
Oil 50.0 ml See Ziehl-Neelsen AFB
Mix well. Label with date and initials,
Working Methylene Blue:
solution is stable for 1 year.
See Ziehl-Neelsen AFB
CAUTION: Flammable.
NOTE: Mineral oil may be substituted for
peanut oil.
Xylene/Peanut Oil Solution:
Xylene 50.0 ml
Peanut Oil 50.0 ml

Mix, solution is stable for 1 year.


CAUTION: Flammable, irritant.

TECH:

DATE:

EXPIRATION:

XYLENE/PEANUT OIL

TECH:

DATE:

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