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>Yes, it does work--you're describing how the Huygens package does it.
>They recommend using beads in the 100-200 nm range and then use a
>theoretical psf to deconvolve the beads and generate an empirical psf.
>Check: http://www.svi.nl or email them at [log in to unmask] .

But this is still a small bead so the signal will be noisy.  What
about using something like a grid pattern?  If the grid dimensions
are known you should, as Peter says, be able to extract the true
PSF (especially in confocal mode where there is no 'missing cone' -
ie no zeros in the OTF.

                                                        Guy

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