Re: FRET on FV500

Posted by Zoltan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/FRET-on-FV500-tp1337622p1338689.html

Hi Julia,

We use PixFRET very successfully, see this reference:

MICROSCOPY RESEARCH AND TECHNIQUE 68:51–58 (2005)

PixFRET, an ImageJ Plug-in for FRET Calculation That Can 

Accommodate Variations in Spectral Bleed-throughs 

JE´ ROˆ ME N. FEIGE,1 DANIEL SAGE,2 WALTER WAHLI,1 BE´ ATRICE DESVERGNE,1 AND LAURENT GELMAN1* 

and AccPbFRET for acceptor photobleaching:

http://www.biophys.dote.hu/accpbfret/

http://www.biomedcentral.com/content/pdf/1471-2105-9-346.pdf

I hope this help,

Zoltan

On Wed, Oct 15, 2008 at 5:56 PM, js1719 <[hidden email]> wrote:

Hello,
 I have a user that is going to do FLIM-FRET on our Olympus FV500 system soon and I was wondering if any other people on the list serve have an established (written) protocol for doing either de-constructive acceptor photo-bleaching (using the protocol processor method) or synthesized emission. We added the FRET module a few years ago but no one here ever used it. He is set up to use CFP/YFP. I have told him to do CFP and YFP alone as controls and I think I will teach him the de-constructive method first since as I remember it being easier. I also wondered what analysis program people use for calculating FRET these days- we tend to use ImageJ a lot for analysis so if you have a plugin that works well can you let me know? I appreciate all the advice you can give me, this is user is completely new to microscopes so the more I can provide him with the better.
Best,
 Julia

--

Zoltan Cseresnyes
Facility manager, Imaging Suite

University of Cambridge, UK