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September 2013

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Subject:
Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?
From:
"Unruh, Jay" <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Fri, 13 Sep 2013 16:44:22 +0000
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Just to balance the discussion, our recent paper in Cell obtained rather different answers than the previously mentioned papers:

http://www.cell.com/abstract/S0092-8674(12)00704-0

If you decide to use the virus particles, I would highly recommend that you control for the GFP quantum yield (the easiest way is with the fluorescence lifetime).

Jay



-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of John Oreopoulos
Sent: Friday, September 13, 2013 11:24 AM
To: [log in to unmask]
Subject: Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?

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Hi Laurent,

Your question reminded me of this impressive piece of work that did the same thing in yeast:

Wu, J.Q. and T.D. Pollard, Counting cytokinesis proteins globally and locally in fission yeast. Science, 2005. 310(5746): p. 310-314.

Wu, J.Q., C.D. McCormick, and T.D. Pollard, Counting proteins in living cells by quantitative fluorescence microscopy with internal standards, in Biophysical tools for biologists, vol 2: In vivo techniques. 2008, Elsevier Academic Press Inc: San Diego. p. 253-+.

Might be of interest to you as well.

John Oreopoulos
Staff Scientist
Spectral Applied Research
Richmond Hill, Ontario
Canada
www.spectral.ca


On 2013-09-13, at 10:34 AM, Gelman, Laurent wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> 
> Hi everybody,
> 
> I somehow remember that I once read an article where viruses expressing/harboring different but known amounts of GFP molecules at their surface where used as a calibration tool for fluorescence quantitative imaging.
> 
> I wish I could retrieve this article but can't find it in Pubmed...
> 
> Thanks in advance for your help!
> 
> Very best regards,
> 
> Laurent.
> 
> _____________________________________________
> Laurent Gelman, PhD
> Head of Facility for Advanced Imaging and Microscopy (Light 
> Microscopy)
> 
> Friedrich Miescher Institut
> WRO 1066.2.16
> Maulbeerstrasse 66
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