Dear confocalists,

(message from a commercial vendor)

The point is, it is not a great effort to compute PSFs with EM
diffraction with today's computers. And they are a sound basis for
comparing HIW values, two-point resolution or other measures in high NA
systems.
> Well, you are right, the convolution with the pinhole is in most cases
> circular symmetric thus simplifying it to a 1D convolution with some
> pre-calculations and the three diffraction integrals are numerically
> easy to manage as well. I suppose the question by the previous poster
For details see:
3D image formation in a high-aperture fluorescence confocal microscope:
a numerical analysis. H.T.M van der Voort and G.J. Brakenhoff,
J.Micr 158 1990, pp. 43-54.
> was not meant to aquire a deconvolution system in order to calculate
> optical sectioning width.
Well, it is not much effort for me to compute PSFs for those who would
like to have one -- oops, not all 1200+ list subscribers...
>
> Anyhow, to what image volume size do the 2.1 seconds refer?
A volume of about 4 x 4 x 8 microns, sampling at Nyquist rate though it
doesn't matter much for the computation time. That is sufficient to
contain a confocal fluorescence PSF with NA=1.3, 488nm 1-photon
excitation, 520nm emission, 250nm backprojected pinhole radius.

Cheers, Hans

--
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dr. Hans T.M. van der Voort                           ([log in to unmask])
Scientific Volume Imaging BV,               URL: http://www.svi.nl/
Alexanderlaan 14, 1213 XS Hilversum,                The Netherlands
tel: ++31-35-6859405, mobile: ++31-653-345445, fax: ++31-35-6837971