Experimental PSFs

[Sarah Kefayati]

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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

 

[jens rietdorf]

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