RE: quest. about fite's AFB

From:

"Johnston, Kathy"

It was my understanding that while it is very important not to over decolorize in any decolorizing solution, the most important step was dewaxing in xylene/oil, to avoid stripping the fat layer, and enhancing staining in the carbol fuchsin. We are using the Fite method in Sheehan, that allows for decolorizing in 1% acid alc. for 1 to 2 minutes. We don't decolorize for even that long, and find 20 to 30 seconds sufficient. However if you are specifically looking for Nocardia with this method, you should decolorize with the sulfuric acid.

Kathy

-----Original Message-----
From: Tony Henwood [mailto:AnthonyH@chw.edu.au]
Sent: Thursday, January 16, 2003 3:47 PM
To: 'Johnston, Kathy'; 'Tague, Curtis'; Histonet (E-mail)
Subject: RE: quest. about fite's AFB

I thought the use of the Fite stain was to demonstrate acid fast, alcohol labile organisms (eg leprosy). Decolourising with acid-alcohol seems very much like a ZN stain, not a Fite.

Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

http://www.histosearch.com/homepages/TonyHenwood/default.html
http://us.geocities.com/tonyhenwoodau/index.html

-----Original Message-----
From: Johnston, Kathy [mailto:Kathy.Johnston@CLS.ab.ca]
Sent: Thursday, 16 January 2003 1:17
To: 'Tague, Curtis'; Histonet (E-mail)
Subject: RE: quest. about fite's AFB

We also use the peanut or mineral oil/xylene to deparaffinize. It helps to
maintain the lipids in the organisms.

In addition we do use 1% acid alcohol (HCl in 70% ETOH) to decolorize, and
have had no problems with overdecolorization in our Fites.

Kathy Johnston
Tech II - Special Stains
Anatomic Pathology - Foothills Medical Center
Calgary Laboratory Services
Ph - 403-944-4760
Fax - 403-270-4093
kathy.johnston@cls.ab.ca

-----Original Message-----
From: Tague, Curtis [mailto:CTague@ahs.llumc.edu]
Sent: Tuesday, January 14, 2003 3:40 PM
To: Histonet (E-mail)
Subject: quest. about fite's AFB

With the peanut oil/xylene step to deparaffinize, is a substitute clearing
agent like histoclear acceptable?

Secondly, just for my knowledge, what purpose does the peanut oil serve, to
simply maintain the structure of the organisms?

Finally, if I didn't have sulfuric acid to differentiate, would acetic acid
or HCl in water work, I figure they're too strong?

Thanks,
curt



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