Re: FITC extracellular quenching

From: bambam (bmalon01@fiu.edu)
Date: Wed Jul 07 1999 - 13:02:32 EST


Robin, I've come across a few articles regarding quenching certain
fluorescence.  Sudan black, trypan blue and percoll (used usually for
density gradients).

Barb

On Tue, 6 Jul 1999, G Robin Barclay wrote:

> 
> Dear colleagues
> Can anyone recommend a method that works for them for quenching FITC-tagged
> agent when bound to the external cell surface but not when the agent is
> internalised by the cell.  Trypan blue and crystal violet come to mind -
> does anyone have this up and running - can you give method/recipes?  I
> assume the quenching agent has to be present in the final cell suspension
> to be run on the cytometer - is it just a matter of resuspending the washed
> FITC-labelled cells in a quencher solution (in PBS?) and running them
> immediately - or are there finite times/temperatures involved.  Is it true
> that only Merck trypan blue works?  Has anyone compared trypan blue to
> crystal violet - and which came out best?  Are there other quenchers which
> may be better?  Do any of these quenchers significantly contaminate the
> cytometer tubing - how do you clean after running?
> Thanks in advance for any help
> Robin Barclay
> SNBTS, Edinburgh
> 



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