FLIM in fixed samples

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Javier Adur Javier Adur
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FLIM in fixed samples

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Dear all.

I am interesting in to know if is valid get FLIM
images from unstained but fixed tissues. Is possible evaluate some biochemical
(FAD or NADH ) information from these images? I already find some works using
FLIM in fixed samples (Conklin et al., 2009 Cell Biochem Biophys DOI
10.1007/s12013-009-9046-7; Tadrous et al., 2003 Journal of Pathology DOI
10.1002/path. 1286, etc.). But I have also read that does not make sense to
work with FLIM in this kind of samples.

 I appreciate any imput about this question.

Thanks in advance.

Javier.





                     
                    Javier F. Adur, Ph.D.
Optics and Photonics Research Center (CEPOF)
Biophotonic Group - IFGW - UNICAMP - BRASIL  
 
                 
yuansheng sun yuansheng sun
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Re: FLIM in fixed samples

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Dear Javier,

As you know, NADH and FAD are metabolic related.  After fixing, you will
probably see little of them.

All the best,
Sheng

Yuansheng Sun, Ph.D.
Keck Center for Cellular Imaging, UVA
http://www.kcci.virginia.edu/workshop/workshop2013/index.php


On Mon, Apr 16, 2012 at 10:31 AM, Javier Adur <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
>
>
>
> Dear all.
>
> I am interesting in to know if is valid get FLIM
> images from unstained but fixed tissues. Is possible evaluate some
> biochemical
> (FAD or NADH ) information from these images? I already find some works
> using
> FLIM in fixed samples (Conklin et al., 2009 Cell Biochem Biophys DOI
> 10.1007/s12013-009-9046-7; Tadrous et al., 2003 Journal of Pathology DOI
> 10.1002/path. 1286, etc.). But I have also read that does not make sense to
> work with FLIM in this kind of samples.
>
>  I appreciate any imput about this question.
>
> Thanks in advance.
>
> Javier.
>
>
>
>
>
>
>                    Javier F. Adur, Ph.D.
> Optics and Photonics Research Center (CEPOF)
> Biophotonic Group - IFGW - UNICAMP - BRASIL
>
>
George McNamara George McNamara
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Re: FLIM in fixed samples

In reply to this post by Javier Adur
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For small molecules - maybe. Do the experiment, publish (in an open
access journal like Alby's latest 3) to let us know.

Fluorescent protein-fluorescent protein heteroFRET : no (thanks Ammasi P).

As for FP-FP homoFRET (fluorescence anisotropy) to measure actin
polymerization - not clear to me, how about a vote by all the FRET
experts here on:

    Vishwasrao HD, Trifilieff P, Kandel ER. In Vivo imaging of the actin
    polymerization state with two-photon fluorescence anisotropy.
    Biophys J. 2012 Mar 7;102(5):1204-14. Epub 2012 Mar 6. PubMed PMID:
    22404943; PubMed Central PMCID: PMC3296026.



Sincerely,

George


On 4/16/2012 10:31 AM, Javier Adur wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
>
>
>
> Dear all.
>
> I am interesting in to know if is valid get FLIM
> images from unstained but fixed tissues. Is possible evaluate some biochemical
> (FAD or NADH ) information from these images? I already find some works using
> FLIM in fixed samples (Conklin et al., 2009 Cell Biochem Biophys DOI
> 10.1007/s12013-009-9046-7; Tadrous et al., 2003 Journal of Pathology DOI
> 10.1002/path. 1286, etc.). But I have also read that does not make sense to
> work with FLIM in this kind of samples.
>
>   I appreciate any imput about this question.
>
> Thanks in advance.
>
> Javier.
>
>
>
>
>
>
>                      Javier F. Adur, Ph.D.
> Optics and Photonics Research Center (CEPOF)
> Biophotonic Group - IFGW - UNICAMP - BRASIL
>
>                  
>