Re: Optical slice thickness and number for PSF and deconvolution

Posted by Guy Cox-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Optical-slice-thickness-and-number-for-PSF-and-deconvolution-tp5415740p5415859.html

This is not calculable on the information you supply.  In principle, if you are prepared to lose resolution in Z you can use fewer sections and a wider pinhole but this does depend on what your software requires.   In the end, the point of doing any sort of deconvolution if you are not sampling at Nyquist is not clear.  IMHO, you would be better off working out what sampling your samples can survive and then dealing with the raw data on those terms. 

 

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Optical Imaging Techniques in Cell Biology

by Guy Cox    CRC Press / Taylor & Francis

     http://www.guycox.com/optical.htm

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Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

 

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From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Jan Trnka
Sent: Thursday, 12 August 2010 8:42 PM
To: [hidden email]
Subject: Optical slice thickness and number for PSF and deconvolution

 

Dear list,

 

this is probably a trivial question but so far I haven't found a good answer. When taking 3D images of subresolution beads in a confocal microscope (for PSF construction) does the number and thickness of slices in the z-stack need to be exactly the same as that of a sample to be deconvolved? I understand the x-y dimensions need to be the same but how does it work for z? Would a higher number of thinner slices (finer z resolution) of the bead improve the construction of the PSF? My actual samples are imaged with a rather wide pinhole setting to limit the exposure of the sample (live cells) and thus provide quite thick optical sections.

 

Thanks,

 

Jan

 

Jan Trnka, MD, PhD

Department of Biochemistry

3rd Medical Faculty

Ruska 87

100 00 Praha 10

Czech Republic

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