Re: Question about deconvolution

Posted by Julio Vazquez on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Question-about-deconvolution-tp7579203p7579205.html

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Hi Christophe,

I won't venture into this discussion other than saying that, in widefield microscopy, contast and resolution are interdependent, and that deconvolution improves both by reducing blur (loss of contrast) due to out of focus signal. There are many good reviews and discussions, including a few chapters in the Confocal Handbook. Another good starting point is here:

http://micro.magnet.fsu.edu/primer/digitalimaging/deconvolution/deconvolutionhome.html

One recent interesting example is work from the Smith lab at Stanford where they use deconvolution on thin sections to effectively achieve super-resolution (~ 100 nm). For example, see:

Wang and Smith,
Sub-diffraction Limit Localization of Proteins in Volumetric Space Using Bayesian Restoration of Fluorescence Images from Ultrathin Specimens.
PLoS Comput Biol. 2012 Aug;8(8):e1002671. Epub 2012 Aug 30.



Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024

http://www.fhcrc.org

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On Oct 23, 2012, at 9:28 AM, Christophe Leterrier wrote:

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>
> Hi folks,
>
> I have a long-standing question regarding deconvolution (as processing
> widefield or confocal images to reassign light from where it originated
> using a PSF).
>
> Is there a theoretical limit to the resolution one could obtain using
> deconvolution? Is is theoretically possible to "break" the diffraction
> limit with deconvolution? That is, to get under the classical 200x200x600nm
> spot? I think it is not the case, but then why would you deconvolve
> widefield or confocal images? What do you gain by doing so on a system that
> is reasonably close to its theoretical capabilities in terms of optical
> performances?
>
> Thanks for your help,
>
> Christophe
>
> --
> Christophe Leterrier
> Researcher
> Axonal Domains Architecture Team
> CRN2M CNRS UMR 7286
> Aix Marseille University, France