Optical slice thickness and number for PSF and deconvolution

Posted by Jan Trnka on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Optical-slice-thickness-and-number-for-PSF-and-deconvolution-tp5415740.html

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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