Posted by
David Baddeley on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Deconvolution-of-3D-SIM-data-tp6251420p6251500.html
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I believe that SIM data is usually at least partly deconvolved already in the
reconstruction process, although the extent of the deconvolution depends on the
heritage of your SIM technique. I think all techniques involve at least a Weiner
filtering step, and some might include a iterative optimisation as well. I'd
thus be pretty cautious about performing any further deconvolution. If you do,
you'd want to use very small beads (<50 nm) for your PSF measurements, which
might make getting reliable (ie not noisy) PSF estimates difficult. Its also
quite likely that the factors which end up limiting the resolution will relate
to uneven bleaching ('burn in' of the mask) and mask errors (alignment errors
and sample abberations which change the shape of the illumination pattern from
what is expected). Deconvolution isn't going to help you here.
cheers,
David
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To:
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Sent: Fri, 8 April, 2011 8:16:08 AM
Subject: Deconvolution of 3D SIM data
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Hi Everyone,
We are starting to process through OMX data sets and we are writing for any
thoughts on the use of deconvolution in these images. We have heard mixed
things about whether or not it is valid... Has anyone here deconvolved 3D SIM
data? Were actual or theoretical PSFs used?
Much thanks,
A