[Histonet] RE: reduction/elimination of red cell autofluorescence on FFPE sections for IF examination

[Amy Porter]

We have been utilizing 0.25% Ammonium Hydroxide(28%) + 70% Ethanol v/v for 1 
hour followed by 10 minutes in 50% Ethanol rinsing in several changes of 
buffer afterward and it seems to be working well for a multitude of 
autofluorescent problems on FFPE sections for us.  We tried to Sodium 
Borohydride and it was a little diffiucult to work with....quite hazardous.

Amy S. Porter, HT (ASCP) QIHC
Investigative HistoPathology Laboratory - Supervisor
2201 Biomedical Physical Sciences Bldg.  Rm #2133
East Lansing, MI  48824-3320
Phone:  (517) 884-5026
Fax:  (517) 432-1368
Email:  portera <@t> msu.edu
Web:  www.humanpathology.msu.edu
----- Original Message ----- 
From: "Thurby, Christina" <christina.thurby <@t> bms.com>
To: "Thurby, Christina" <christina.thurby <@t> bms.com>; 
<histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, August 24, 2009 11:00 AM
Subject: [Histonet] RE: reduction/elimination of red cell autofluorescence 
on FFPE sections for IF examination


Sorry I did not include my contact information.
Christina Thurby
christina.thurby <@t> bms.com
812-429-8097
Thanks!

>-----Original Message-----
>From: Thurby, Christina
>Sent: Monday, August 24, 2009 9:59 AM
>To: 'histonet <@t> lists.utsouthwestern.edu'
>Subject: reduction/elimination of red cell autofluorescence on FFPE 
>sections for IF
>examination
>
>Can anyone give feedback on reagents/procedures to use for 
>immunofluorescence to
>reduce/eliminate red cell autofluorescence on FFPE sections.  I am using an 
>indirect method
>of labeling for two antibodies (FITC and Texas Red).
>
>I have not tried 0.1% sodium borohydride.  Will this help for formalin 
>fixed specimens?  I
>have read that it is used for gluteraldehyde autofluorescence reduction. 
>If appropriate how
>long should this reagent be applied to the specimen and at what 
>temperature.

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