"Johnston, Kathy" wrote:

 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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