Confocal Microscopy List

Re: FRET results

Posted by Periasamy, Ammasi (ap3t) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/FRET-results-tp2580297p2580759.html

Usually we use 514 nm line from the argon laser to bleach or excite the YFP or Venus. 488nm is close to CFP excitation wavelength and you may bleach a lot of CFP molecule compared with 514nm.

Ammasi Periasamy, Ph.D.

Director, Keck Center for Cellular Imaging (KCCI)

Professor of Biology and Biomedical Engineering

Biology, Gilmer Hall (064), McCormick Rd

University of Virginia

Charlottesville, VA 22904

Voice: 434-243-7602 (Office); 982-4869 (lab)

Fax:434-982-5210; Email:[hidden email]

http//:www.kcci.virginia.edu

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Workshop on FRET Microscopy, March 9-13, 2010

http://www.kcci.virginia.edu/workshop/workshop2010/index.php

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From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of ramesh kandpal
Sent: Friday, April 03, 2009 7:56 AM
To: [hidden email]
Subject: FRET results

hi,

can anyone give some idea that which is the most appropriate laser wavelength for performing acceptor photobleaching method of CFP/YFP FRET. If 488nm laser is used for YFP excitation is there any possibility of loosing FRET data. What difference does it create in FRET results if we use 514nm laser line.

thanks & best wishes,