FLIM/FRET imaging - achievements

[Alessandro Esposito]

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Dear all,

   once in a while I am asked if there was any biological discovery that was possible only because of FLIM.

Even though time-resolved imaging is my bread and butter, I have difficulties to enumerate the top achievements in terms of biological discoveries for such technologies. Could you share your opinion about the top discoveries in biology where FLIM (and perhaps FRET imaging in general) had a critical role?


Cheers,

Alessandro

[John Oreopoulos]

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Allesandro,

I recommend taking a look at the recent work of Enrico Gratton's Group  
at the LFD, and in particular the phasor approach for FLIM. I think  
there were a few review papers from this group that also discussed new  
information gained about certain biological processes that was only  
possible with phasors and FLIM-FRET.

Sincerely,

John Oreopoulos


Quoting Alessandro <[hidden email]>:

> *****
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> *****
>
> Dear all,
>
>    once in a while I am asked if there was any biological discovery  
> that was possible only because of FLIM.
>
> Even though time-resolved imaging is my bread and butter, I have  
> difficulties to enumerate the top achievements in terms of  
> biological discoveries for such technologies. Could you share your  
> opinion about the top discoveries in biology where FLIM (and perhaps  
>  FRET imaging in general) had a critical role?
>
>
> Cheers,
>
> Alessandro
>

[Tim Feinstein]

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Regarding FRET, you certainly have a rich field to draw on.  A sizable fraction of all genetically encoded biosensors use FRET.   Protein-protein interaction studies have overall benefited greatly from FRET out. If you include BRET, Michel Bouvier’s work (for example) has made an enormous impact on the field of GPCR pharmacology.  
               
I have seen FLIM contribute most significantly to receptor tyrosine kinase dimerization studies, maybe best exemplified in the work of Phillippe Bastiaens.  Though I do not recall the exact reference I remember a breakthrough FLIM study from maybe 15 (?) years ago that showed a wave of transient RTK dimerization propagating from the point of contact between a cell and a bead coated with RTK agonist.  I think the unique qualities of RTKs – relatively transient interactions against a large background of non-interacting proteins – makes them uniquely suitable for FLIM.  

Best,


Tim

Timothy Feinstein, Ph.D.
Research Scientist
University of Pittsburgh Department of Developmental Biology


On 6/6/17, 12:45 PM, "Confocal Microscopy List on behalf of Alessandro" <[hidden email] on behalf of [hidden email]> wrote:

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    *****
   
    Dear all,
   
       once in a while I am asked if there was any biological discovery that was possible only because of FLIM.
   
    Even though time-resolved imaging is my bread and butter, I have difficulties to enumerate the top achievements in terms of biological discoveries for such technologies. Could you share your opinion about the top discoveries in biology where FLIM (and perhaps FRET imaging in general) had a critical role?
   
   
    Cheers,
   
    Alessandro
   

[Michelle Digman]

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Dear Alessandro,

Yes there are many examples. From understanding Cdc42, Rac1 and Rho GTPAse
to ion transport proteins. These are all biologically important Here are a
few to start off with:

Herrington KA, Trinh AL, Dang C, O’Shaughnessy E, Hahn KM, Gratton E,
Digman MA, Sütterlin C.
Spatial analysis of Cdc42 activity reveals a role for plasma
membrane-associated Cdc42 in centrosome regulation. Mol Biol Cell. 2017;
[Epub ahead of print].

Digman MA Gratton E.
The phasor approach to fluorescence lifetime imaging: exploiting phasor
linear properties.
Fluorescence Lifetime Spectroscopy and Imaging: Principles and Applications
in Biomedical Diagnostics. By L Marcu, PMW French, and DS Elson (Editors).
CRC Press, pp. 235-248, 2014. ISBN: 9781439861677

Hinde E, Yokomori K, Gaus K, Hahn KM, Gratton E.
Fluctuation-based imaging of nuclear Rac1 activation by protein
oligomerisation.
Sci Rep. 2014; 4, 4219. PMCID: PMC3936235

Hinde E, Digman MA, Hahn KM, Gratton E.
Millisecond spatiotemporal dynamics of FRET biosensors by the pair
correlation function and the phasor approach to FLIM.Proc Natl Acad Sci
USA. 2013; 110(1): 135-140. PMCID: PMC3538204

 Dobrinskikh E, Lanzano L, Rachelson J, Cranston D, Moldovan R, Lei T,
Gratton E, Doctor RB.
Shank2 contributes to the apical retention and intracellular redistribution
of NaPiIIa in OK cells.
Am J Physiol Cell Physiol. 2013; 304(6): C561-573. PMCID: PMC3674434

Giral H, Cranston D, Lanzanò L, Caldas Y, Sutherland E, Rachelson J,
Dobrinskikh E, Weinman EJ, Doctor RB, Gratton E, Levi M. NHE3 regulatory
factor 1 (NHERF1) modulates intestinal Na-dependent phosphate transporter
(NaPi-2b) expression in apical microvilli. J Biol Chem. 2012; 287(42):
35047-35056. PMCID: PMC3471691

Hinde E, Digman MA, Welch C, Hahn KM, Gratton E.
Biosensor Förster resonance energy transfer detection by the phasor
approach to fluorescence lifetime imaging microscopy. Microsc Res Tech.
2012; 75(3): 271-281. PMCID: PMC352310

Digman MA, Caiolfa VR, Zamai M, Gratton E.
The Phasor approach to fluorescence lifetime imaging analysis.
Biophys J. 2008; 94(2): L14-16. PMCID: PMC2157251


*******************************
Michelle A. Digman
Assistant Professor
Department of Biomedical Engineering
University of California Irvine
<[hidden email]>

*http://faculty.sites.uci.edu/digmanlab/
<http://faculty.sites.uci.edu/digmanlab/>*
email: [hidden email]
phone: [phone]
fax: [phone]


On Tue, Jun 6, 2017 at 9:45 AM, Alessandro <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all,
>
>    once in a while I am asked if there was any biological discovery that
> was possible only because of FLIM.
>
> Even though time-resolved imaging is my bread and butter, I have
> difficulties to enumerate the top achievements in terms of biological
> discoveries for such technologies. Could you share your opinion about the
> top discoveries in biology where FLIM (and perhaps FRET imaging in general)
> had a critical role?
>
>
> Cheers,
>
> Alessandro
>

[Andrew-2]

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Alessandro,

I came across this rather interesting FRET paper some time ago, they used
it to create a tension sensor to get a measure on how much force was being
applied through a protein.

Fluid shear stress on endothelial cells modulates mechanical tension across
VE-cadherin and PECAM-1.
Conway DE, Breckenridge MT, Hinde E, Gratton E, Chen CS, Schwartz MA.
Curr Biol. 2013 Jun 3;23(11):1024-30. doi: 10.1016/j.cub.2013.04.049. Epub
2013 May 16.
PMID: 23684974


Andrew

On Tue, Jun 6, 2017 at 12:45 PM, Alessandro <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all,
>
>    once in a while I am asked if there was any biological discovery that
> was possible only because of FLIM.
>
> Even though time-resolved imaging is my bread and butter, I have
> difficulties to enumerate the top achievements in terms of biological
> discoveries for such technologies. Could you share your opinion about the
> top discoveries in biology where FLIM (and perhaps FRET imaging in general)
> had a critical role?
>
>
> Cheers,
>
> Alessandro
>

[Alessandro Esposito]

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Dear John,

    thank you for the reply, but I was actually looking after something different. I am looking for the discoveries with the highest impact (not IF) to biology that could not have been achieved without FLIM. I am fully aware of how useful FLIM, phasors, and Enrico's huge contribution to the field, including the Michelle's contribution (to whom I'll reply in a moment).

Let's say we would like to compile a list of 10 papers, primary research describing biological discoveries of high signifance where FLIM had an essential role.

Cheers,

Alessandro
www.quantitative-microscopy.org

[Alessandro Esposito]

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Dear Tim,

   thank you for your comments. I am actually curious to follow up citations within certain papers to see how certain works have contributed to shape the field. Perhaps, I'll write up a page to publish on a blog and submit this first here for comments.

Cheers,

Alessandro
www.quantitative-microscopy.org

[Alessandro Esposito]

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Dear Michelle,

   thank you for the list. Of course, I am aware about the methodological developments and their usefulness. I was not aware, however, of some of the most recent biological applications. Amongst others, the Cdc42 paper is clearly a very good example. It is not trivial for me to select the most topical biological discoveries where FLIM played a fundamental role.

Cheers,

Alessandro
www.quantitative-microscopy.org

[Alessandro Esposito]

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Dear Andrew,

  thank you for the reference, I will add this to my list. For those works outside my area of expertise, I have more difficulties to assess the potential impact of the discovery, but it seems a ratehr good example.

Cheers,

Alessandro
www.quantitative-microscopy.org

[Romain Laine]

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Hi Alessandro,
If you want to venture on the side of the medical applications of FLIM, you
may want to have a look at the work of Laura Marcu from UC Davis.
http://bme.ucdavis.edu/people/departmental-faculty/laura-marcu-ph-d/

She is pushing the effort towards translation of the technology for
clinical applications, label-free, etc.
Cheers,

Romain

On Wed, Jun 7, 2017 at 10:30 AM, Alessandro <
[hidden email]> wrote:

> *****
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> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Andrew,
>
>   thank you for the reference, I will add this to my list. For those works
> outside my area of expertise, I have more difficulties to assess the
> potential impact of the discovery, but it seems a ratehr good example.
>
> Cheers,
>
> Alessandro
> www.quantitative-microscopy.org
>