Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Suresh,
Did you do the Sudan Black B treatment before or after the applying  
the secondary antibodies?

regards,
Glen


Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
(206) 616-4156
[log in to unmask]

************************************************************************ 
******
The box said "Requires Windows 95 or better", so I bought a Macintosh.
************************************************************************ 
******


On Nov 30, 2006, at 3:51 AM, suresh mehta wrote:

> Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi- 
> bin/wa?S1=confocal
> Hi everybody
> I have to work on brain tissue particularly fresh frozen  
> cryosection for immunoflourecence. I am novice to this field  
> however i have previously done some experiment where i got some  
> problem with probably from autoflourecence molecules (lipofuschin).  
> I have used Sudan Black B to reduce the autoflourecence but instead  
> fails to detect the specific signal. I have used secondary antibody  
> conjugated with FITC or texus Red primarily raised in donkey. I  
> could see some signal of FITC but when double labelled with Texus  
> red, I failed to see any positive signal of Texus red after Sudan  
> Black B treatment.
>
> Now I have to restart the experimentation and purchased Alexa dyes  
> (488 & 594 molecular probes) labelled secondary antibody to ensure  
> that I may not miss the signal may be due to photobleaching.
> So my request to all user is
> 1) can you guide me regarding the processing of cryosection for  
> immunoflourescence
> 2) Can you provide me the protocol for double lablling of protein  
> on same section (Cryosection)
> 2) Any suggestion to restart my work
> 3) Any stringent precaution to be taken during experiment..........
>
> With thanks in anticipation and regards
>
> Suresh Mehta
> Division of Pharmacology
> central Drug Research Institute
> Lucknow India
>
> Find out what India is talking about on - Yahoo! Answers India
> Send FREE SMS to your friend's mobile from Yahoo! Messenger Version  
> 8. Get it NOW