Dear Sheng,

I also have used Cy3 as a donor in experiment before. However I read it 
has a short lifetime compared to GFP for example. And so I was wondering 
if anyone was successful using it as a donor in a FLIM experiment.

Thank you 

JP



 

- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Jean-Pierre CLAMME, PhD
Chief Scientist
Nitto Denko Technical
501 Via Del Monte
Oceanside, CA 92058
E-mail: [log in to unmask]
Phone: +760.435.7065


Confocal Microscopy List <[log in to unmask]> wrote on 
03/20/2012 09:59:04 AM:

> yuansheng sun <[log in to unmask]> 
> Sent by: Confocal Microscopy List <[log in to unmask]>
> 
> 03/20/2012 09:59 AM
> 
> Please respond to
> Confocal Microscopy List <[log in to unmask]>
> 
> To
> 
> [log in to unmask]
> 
> cc
> 
> Subject
> 
> Re: FLIM using Cy3 as a donor ?
> 
> *****
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> Dear Jean-Pierre,

> I guess you are trying to do FLIM-FRET with Cy3 as the FRET donor.  I 
have
> not personally used Cy3 as the FRET donor.  But I think Cy3 can be a 
FRET
> donor, depending upon what is the acceptor.  There is a paper (given 
below)
> from Taekjip Ha group, where they used Cy3 as the FRET donor.

> Single-Molecule Three-Color FRET.  Biophys J. 2004 August; 87(2): 
1328–1337

> There are other considerations - Would you use 1p or 2p for FLIM?  If 
you
> will use 2p, you need to make sure that you can excite Cy3 efficiently 
with
> your 2p laser.  In FLIM-FRET analysis, you want your donor-alone (Cy3)
> decay to be simple - ideally mono-exponential.  It is worth searching if
> anybody has measured the lifetime of Cy3, since you do not have a system 
on
> site.  However, it is always important to know what is the lifetime of 
Cy3
> in your sample.

> I hope it helps.  And good luck.

> Sheng
> Yuansheng Sun, Ph.D.
> Keck Center for Cellular Imaging
> University of Virginia

> We held a workshop on FRET Microscopy
> http://www.kcci.virginia.edu/workshop/workshop2013/index.php

> 
> On Mon, Mar 19, 2012 at 8:47 PM, Jean-Pierre CLAMME <
> [log in to unmask]> wrote:

> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hi,
> >
> > I was wondering if anyone had done or tried FLIM experiments using Cy3 
(or
> > Dy547)  as a donor. I have seen a few FLIM publications were Cy3 is 
the
> > acceptor for GFP but none were Cy3 is the donor.
> > I don't have a FLIM system handy so I can't just go a try it. So I'm
> > looking for this information to determine if what I want to do is 
feasible
> > before I go and find a Lab were I could do the experiment.
> >
> > Thank you in advance
> >
> > JP
> >
> > - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
> > Jean-Pierre CLAMME, PhD
> > Chief Scientist
> > Nitto Denko Technical
> > 501 Via Del Monte
> > Oceanside, CA 92058
> > E-mail: [log in to unmask]
> > Phone: +760.435.7065
> >