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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