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Ricardo,

one could think that 2D processing is a special case of 3D processing (with
Z=1), but I have to raise some caution. Any good 3D deconvolution package
has implemented some kind of internal data expansion to avoid the
circularity inherent to the FFT convolutions, which would cause edge
artefacts when not done. Going into such algorithm with 2D data will
internally produce a 3D data set, which may, depending on the
implementation, not be optimal for solving your problem. Taking the PSF at
the in-focus position for 2D deconvolution is correct.

Generally, the 2D restoration effect will not become as much apparent as in
the 3D case, unless you plan to do it on an othogonal x-z or y-z section
(with appropriate orthogonal PSF section). This is because you do not
account for the contributions from other focal sections. The PSF has the
greatest (in fact infinite) region of support in axial direction, while in
transverse direction its approximately only the size of the Airy disk. So I
hope you do not expect too much ...

Cheers
Lutz




----- Original Message ----- 
From: "Ricardo Armisen" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Thursday, March 23, 2006 1:58 AM
Subject: home made deconvolution setup


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi all
> a question about deconvolution (again!!!!!)
> I am trying to put together a "home made" time lapse microscope (for gfp
> tag proteins and slow calcium signals) with deconvolution in mind,
> would something like this work?
>
> -a IM 35 zeiss inverted microscope (the 160 length tube!!) with a good
> 63x/NA 1.3 oil objective (but no luck getting a water one so far)
> - a spot 2 (slider) ccd camera (diagnostic instruments)
> - a jena piezo device for the objective (any bet if the extra 11 mm will
> interfere with the optics?)..this is optional
> - a shutter for the mercury lamp and an optical excitation switch
(DX-1000,
> Solameretech)
> - run by and old version of QED imaging software
>
>  the deconvolution would be made afterwards image acquisition. I don't
plan
> to do any fancy 4D imaging, just one picture (one focal plane) over time,
> but most deconvolution free software (ImageJ and Image Track) I had found
> seem to be done for xyz stacks, I was able to trick the software supplying
> a psf made  of only one picture (not much of a psf...), but I am not sure
> if this is the rigth procedure, do you think it is posible to get good
> plain 2d deconvolution?  thanks....
>
> Ricardo Armisen
>
> SUNY at SB
>
> U. de Chile