Nature Methods article: YFP>CFP photoconversion

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3 total messages Started by Stephan Lange Tue, 08 Nov 2005 21:56

#65

Author: Stephan Lange
Date: Tue, 08 Nov 2005 21:56

17 lines
1668 bytes

Hello,

  anyone came accross the phenomenon that YFP emission spectra change 
into CFP-like emission spectra upon photobleaching?

see "Photoconversion of YFP into a CFP-like species during acceptor 
photobleaching FRET experiments"Guillaume Valentin, C�line Verheggen, 
Tristan Piolot, Henry Neel, Ma�t� Coppey-Moisan & Edouard Bertrand 
doi:10.1038/nmeth1105-801; 
http://www.nature.com/nmeth/journal/v2/n11/index.html)

A friend of mine tried it today with pEYFP (Clontech) in tranfected and 
PFA fixed Cos-1 cells and could not reproduce the reported effect...?!
Any help or experience in that case would be highly appreciated...

kind Regards
Stephan

Re: Nature Methods article: YFP>CFP photoconversion

#66

Author: rene
Date: Thu, 10 Nov 2005 15:18

27 lines
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Couldn't reach the article, nor do I know ins and outs of fret, but 2
things come to mind: if yfp changes to cfp, this would imply changes on
molecular level, therefore 1) is fixation allowed, and 2) would it not
be important to use exactly the right yfp??

Just a thought, Rene.


Stephan Lange schreef:

> Hello,
>
>   anyone came accross the phenomenon that YFP emission spectra change
> into CFP-like emission spectra upon photobleaching?
>
> see "Photoconversion of YFP into a CFP-like species during acceptor
> photobleaching FRET experiments"Guillaume Valentin, Céline Verheggen,
> Tristan Piolot, Henry Neel, Maïté Coppey-Moisan & Edouard Bertrand
> doi:10.1038/nmeth1105-801;
> http://www.nature.com/nmeth/journal/v2/n11/index.html)
>
> A friend of mine tried it today with pEYFP (Clontech) in tranfected and
> PFA fixed Cos-1 cells and could not reproduce the reported effect...?!
> Any help or experience in that case would be highly appreciated...
> 
> kind Regards
> Stephan

Re: Nature Methods article: YFP>CFP photoconversion

#67

Author: Stephan Lange
Date: Thu, 10 Nov 2005 16:22

21 lines
1920 bytes

Yeah, Nature requires that your institution has a special subscription 
to view the new arcticles... (no comment about that; grrrr)

fixation is allowed. They have shown the effect on fixed cells as well 
as YFP coupled to beads (non fixed)

They used also different types of YFP. The one from Clontech as well as 
other YFP variants (see supplementary data). All investigated YFP-types 
showed this "phenomenon"...

rene wrote:

> Couldn't reach the article, nor do I know ins and outs of fret, but 2
> things come to mind: if yfp changes to cfp, this would imply changes on
> molecular level, therefore 1) is fixation allowed, and 2) would it not
> be important to use exactly the right yfp??
> 
> Just a thought, Rene.
> 
>

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