[Histonet] Quenching auto fluorescence in human tissue

[WILLIAM DESALVO]

I have not used "Chicago Blue" (Inoticed the product is discontinued by Sigma Aldrich) for quenching, but have often used "Evans Blue" (synonym: Direct Blue 53; Empirical Formula: C34H24N6Na4O14S4 ;Molecular Weight: 960.81;CAS Number: 314-13-6) without any issue. Sigma Aldrich sells the Evans Blue is solution (>75% dye content). You might want to give the Evans Blue a try (product # E2129).

William DeSalvo, B.S., HTL(ASCP)

 

> Date: Wed, 18 Jan 2012 16:11:51 +1100
> From: daniela.bodemer <@t> mcri.edu.au
> To: Histonet <@t> lists.utsouthwestern.edu; histonet-request <@t> lists.utsouthwestern.edu
> CC: 
> Subject: [Histonet] Quenching auto fluorescence in human tissue
> 
> Hi all,
> 
> 
> 
> I have been experiencing trouble with my secondary antibody when doing
> my immuno with human colon tissue.
> 
> I use to use FITC 488 as my secondary antibody and changed over to
> Alexa488 due to the FITC's manufacturing stop.
> 
> The auto fluorescence of the human tissue is quenched with Chicago Blue
> solution and the slides are mounted with Prolong Gold antifade reagent
> to extend the life of the immuno slides.
> 
> The trouble is, after a few days the fluorofors seem to 'detach' and
> start floating around, not allowing reliable validation of the results.
> This also happen if I use Mowiol as a mounting medium.
> 
> Is the mounting medium reacting with Chicago Blue? Is there any
> alternative to quench die auto fluorescence of the tissue?
> 
> 
> 
> Thanks for your ideas.
> 
> 
> 
> Daniela Bodemer 
> 
> Research Assistant
> 
> Surgical Research, Infection and Immunity
> 
> 
> 
> Murdoch Childrens Research Institute
> 
> The Royal Children's Hospital
> 
> Flemington Road Parkville Victoria 3052 Australia 
> 
> T 03 9936 6676 T (03 9936 6794) 
> 
> E daniela.bodemer <@t> mcri.edu.au <mailto:firstname.surname <@t> mcri.edu.au> 
> 
> www.mcri.edu.au <http://www.mcri.edu.au/> 
> 
> 
> 
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