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February 2008

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Subject:
From:
Jens Rietdorf <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Thu, 28 Feb 2008 11:24:33 -0800
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Sarah,

you may use a theoretical PSF, for which you have to enter a number of
parameters including sample RI. If you want to use an experimental PSF,
Huygens Pro uses a number of averaged bead images to back-calculate the PSF,
for this, you need to give in addition the radius of the beads used.

In any case, I strongly recommend to avoid distortions of the PSF by RI
mismatch, which will, deco or not, reduce the resolution of the images.
Deconvolution can only compensate for very moderate distortions of the PSF.
Try to compensate for asymmetric PSF by adapting the immersion fluid RI.

cheers, jens


Sarah Kefayati wrote:
> 
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> 
> Hello all!
> 
> I have couple of questions and I rally appreciate if you could help me:
> 
> 
> Is theoretical PSF the first data to deconvolve the Experimental PSFs for
> the deconvolution algorithms?
> 
> If so, for a Huygens Professional software, is necessary to provide the
> sample refractive index for the program to calculate Theoretical PSF?(I am
> collecting in situ PSFs from a complex sample)
> 
> For Huygens pro,is that an option to define a PSF and ask the software to
> adapt that and operate the deconvolution with that PSF?
> 
> Thank you
> Sarah
> 
> 

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