FRET calculations

[Boswell, Carl A - (cboswell)]

To whomever contacted me regarding spectral data for FRET calculations:  I've managed to lose your message, and hence email contact, within 24 hrs of receiving it.  Can you please resend?

Where does this stuff go, anyway, to the Vortex of Single Socks?

Thanks,
C


Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
Univ. of Arizona
520-954-7053
FAX [phone]

[Craig Brideau]

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Bit rot is the usual culprit. @:-)

Craig



On Wed, Apr 27, 2011 at 5:05 PM, Boswell, Carl A - (cboswell) <
[hidden email]> wrote:

> To whomever contacted me regarding spectral data for FRET calculations:
>  I've managed to lose your message, and hence email contact, within 24 hrs
> of receiving it.  Can you please resend?
>
> Where does this stuff go, anyway, to the Vortex of Single Socks?
>
> Thanks,
> C
>
>
> Carl A. Boswell, Ph.D.
> Molecular and Cellular Biology
> Univ. of Arizona
> 520-954-7053
> FAX [phone]
>
>
>

[Christian-103]

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I have a user who is trying to express CFP, YFP, mCherry and mPlum in the same cells.  I have not had mPlum in my hands, but notice it has roughly the same excitation as mCherry.  Without using the spectral detector, if the expression levels are similar, should I expect these two to be a problem?  Thanks.

[Periasamy, Ammasi (ap3t)]

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Excitation wavelength for mCherry and mPlum is the same 590 nm but the emission for mCherry is 610nm and for mPlum is 650 nm. There is about 40 nm separation but it may be a problem if you are trying for any colocalization investigation. If you can use a narrow band emission filters combined with high sensitivity detectors (APD or HD), you can separate the two images with one excitation wavelength. There is always another question whether the cell can express well with so many fluorophore.

Dr. Ammasi Periasamy
Professor & Center Director
Keck Center for Cellular Imaging (KCCI)
Biology, Gilmer Hall (064), 485 McCormick Rd
University of Virginia
Charlottesville, VA 22904
(Campus Mail - P.O. Box 400328)
Voice: 434-243-7602 (Office); 982-4869 (lab)
Fax:[phone]; Email:[hidden email]
http://www.kcci.virginia.edu/Contact/peri.php 
************************
11th Annual Workshop on FRET Microscopy, March 5-10, 2012
http://www.kcci.virginia.edu/workshop/workshop2012/ 
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-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christian
Sent: Friday, August 19, 2011 2:01 PM
To: [hidden email]
Subject: mCherry and mPlum

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To join, leave or search the confocal microscopy listserv, go to:
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I have a user who is trying to express CFP, YFP, mCherry and mPlum in the same cells.  I have not had mPlum in my hands, but notice it has roughly the same excitation as mCherry.  Without using the spectral detector, if the expression levels are similar, should I expect these two to be a problem?  Thanks.