Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
mTFP (mTFP1) to mVenus or mKO looks good. mTFP is
a better match for a 457nm laser line than
Cerulean and does not have some of Cerulean's
funky behavior (described in another article in
the JBO issue below). For mTFP -> Venus see:
http://spiedl.aip.org/getabs/servlet/GetabsServlet?prog=normal&id=JBOPFO000013000003031203000001&idtype=cvips&gifs=Yes
<http://spiedl.aip.org/journals/doc/SPIEDL-home/jrnls/>J.
Biomed. Opt. /
<http://spiedl.aip.org/getabs/servlet//dbt/dbt.jsp?KEY=JBOPFO&Volume=13>Volume
13 /
<http://spiedl.aip.org/getabs/servlet//dbt/dbt.jsp?KEY=JBOPFO&Volume=13&Issue=3>Issue
3 /
<http://spiedl.aip.org/dbt/dbt.jsp?KEY=JBOPFO&Volume=13&Issue=3#Special
Section on Visible Fluorescent Proteins>Special
Section on Visible Fluorescent Proteins
Characterization of an improved donor fluorescent
protein for Förster resonance energy transfer microscopy
J. Biomed. Opt., Vol. 13, 031203 (2008); DOI:10.1117/1.2939094
<http://spiedl.aip.org/getabs/servlet//vsearch/servlet/VerityServlet?KEY=ALL&possible1=Day%2C+Richard+N.&possible1zone=author&maxdisp=25&smode=strresults&aqs=true>Richard
N. Day and
<http://spiedl.aip.org/getabs/servlet//vsearch/servlet/VerityServlet?KEY=ALL&possible1=Booker%2C+Cynthia+F.&possible1zone=author&maxdisp=25&smode=strresults&aqs=true>Cynthia
F. Booker
University of Virginia Health System, Departments
of Medicine and Cell Biology, P. O. Box 800578,
Charlottesville, Virginia 22908-0578
<http://spiedl.aip.org/getabs/servlet//vsearch/servlet/VerityServlet?KEY=ALL&possible1=Periasamy%2C+Ammasi&possible1zone=author&maxdisp=25&smode=strresults&aqs=true>Ammasi
Periasamy
University of Virginia, W.M. Keck Center for
Cellular Imaging, Departments of Biology and
Biomedical Engineering, Gilmer Hall, Charlottesville, Virginia 22904-4328
The genetically encoded fluorescent proteins
(FP), used in combination with Förster resonance
energy transfer (FRET) microscopy, provide the
tools necessary for the direct visualization of
protein interactions inside living cells.
Typically, the Cerulean and Venus variants of the
cyan and yellow FPs are used for FRET studies,
but there are limitations to their use. Here,
Cerulean and the newly developed monomeric Teal
FP (mTFP) are compared as FRET donors for Venus
using spectral and fluorescence lifetime
measurements from living cells. The results
demonstrate that when compared to Cerulean, mTFP
has increased brightness, optimal excitation
using the standard 458-nm laser line, increased
photostability, and improved spectral overlap
with Venus. In addition, the two-photon
excitation and fluorescence lifetime
characteristics are determined for mTFP.
Together, these measurements indicate that mTFP
is an excellent donor fluorophore for FRET
studies, and that its use may improve the
detection of interactions involving proteins that
are difficult to express, or that need to be produced at low levels in cells.
<http://www.ncbi.nlm.nih.gov/sites//pubmed/18325109?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum>Ai
HW, Olenych SG, Wong P, Davidson MW, Campbell RE.
<http://www.ncbi.nlm.nih.gov/sites//pubmed/18325109?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum>
Free in PMC
Hue-shifted monomeric variants of Clavularia cyan
fluorescent protein: identification of the
molecular determinants of color and applications in fluorescence imaging.
BMC Biol. 2008 Mar 6;6:13.
PMID: 18325109 [PubMed - indexed for MEDLINE]
<http://www.ncbi.nlm.nih.gov/sites//pubmed/16859491?ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum>Ai
HW, Henderson JN, Remington SJ, Campbell RE.
Directed evolution of a monomeric, bright and
photostable version of Clavularia cyan
fluorescent protein: structural characterization
and applications in fluorescence imaging.
Biochem J. 2006 Dec 15;400(3):531-40.
PMID: 16859491
At 09:53 AM 7/10/2008, yee z wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi Everyone,
>
>
>
>I start a new job in the imaging core recently,
>and think to give a demo show on how fret works
>in our new live cell imaging system.
>
>
>
>I would highly appreciate if anyone on this list
>can provide me an easy working FRET pair like
>plasmids that I can transfect cell with or let
>me know where I can purchase such things.
>
>
>
>
>
>PS: I am located in Boston, USA.
>
>
>
>Thanks,
>
>
>
>Yi
>
>
>
>
George McNamara, Ph.D.
University of Miami, Miller School of Medicine
Image Core
Miami, FL 33010
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[log in to unmask]
305-243-8436 office
http://home.earthlink.net/~pubspectra/
http://home.earthlink.net/~geomcnamara/
http://www.sylvester.org/research/SR_lab_analytical.asp?ana=desc
(Analytical Imaging Core Facility)
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