Confocal Microscopy List

Quantifying GFP intensity with GFP-decorated viruses as a reference?

classic Classic list List threaded Threaded
♦
♦
 5 messages Options
lgelman lgelman
Reply | Threaded
Open this post in threaded view
♦
♦
|

Quantifying GFP intensity with GFP-decorated viruses as a reference?

*****
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
CH-4058 Basel
+41 (0)61 696 43 38
+41 (0)79 618 73 69
Jolien Tyler Jolien Tyler
Reply | Threaded
Open this post in threaded view
♦
♦
|

Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

One recent paper that used viral particles as a standard for quantitative microscopy is Lawrimore, Bloom and Salmon, 2011, JCB (http://www.ncbi.nlm.nih.gov/m/pubmed/22084307/)
I don't know that they use multiple different viral particles, but it may be a place to start.

Hope that helps,

Jolien Verdaasdonk

Bloom Lab
UNC Chapel Hill


On Sep 13, 2013, at 10:34 AM, "Gelman, Laurent" <[hidden email]> 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
> CH-4058 Basel
> +41 (0)61 696 43 38
> +41 (0)79 618 73 69
berkserk berkserk
Reply | Threaded
Open this post in threaded view
♦
♦
|

Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?

In reply to this post by lgelman
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Laurent,

Maybe this would help, http://www.jbc.org/content/276/31/29361.full .

Thanks
Serkan Berk

University of Minnesota,
116 Church St. SE
Minneapolis, MN 55455
612-6246045
John Oreopoulos John Oreopoulos
Reply | Threaded
Open this post in threaded view
♦
♦
|

Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?

In reply to this post by lgelman
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

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
> CH-4058 Basel
> +41 (0)61 696 43 38
> +41 (0)79 618 73 69
Unruh, Jay Unruh, Jay
Reply | Threaded
Open this post in threaded view
♦
♦
|

Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

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:[hidden email]] On Behalf Of John Oreopoulos
Sent: Friday, September 13, 2013 11:24 AM
To: [hidden email]
Subject: Re: Quantifying GFP intensity with GFP-decorated viruses as a reference?

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

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
> CH-4058 Basel
> +41 (0)61 696 43 38
> +41 (0)79 618 73 69
Free forum by Nabble Edit this page