I am aware that I could use deconvolution algorithms such as LUCY, Weiner and Blind but I am confused as to how to estimate the PSF (point spread function), which is needed for the LUCY and Weiner algorithms. I am currently using the Widefield Florescence Microscopy setup.
I have read in a lot of places around the internet that a fluorescent bead that represents a sub resolution object and therefore a impulse function. While I understand that the OTF (optical transfer function) is represented by the image I am getting, how am I superposed to deconvolve the image I have using the very same image?
Here is an example of the image I am trying to deconvolve:
I have a program which automatically singles in on the bead and crops the rest of the image. All I need to do is apply deconvolution to this.
My ultimate aim to achieve optical sectioning by deconvolving a bunch of 2D slices I got from a video and representing them with a 3D software.
So can someone please explain what I need to do, either theoretically (or in terms of MATLAB, which would be preferred).
One Possible Solution
After talking to a few people, I have noticed that EPFL have a Java resource that is callable from MATLAB. It's a PSF generator, based on the specifications of your microscope. This generated PSF can be used with the LUCY and Weiner.
I have also found this resource, which is a MATLAB library dedicated to what I am working on right now.
This (I would love it, if someone attempts to answer this question too) is how I will measure the performance of the deconvolution algorithms. I believe that the ultimate result of my deconvolution will depend on the PSF estimation (EPFL resource or Praveen's algorithm) and the deconvolution algorithm (Blind, LUCY, Weiner etc.). I will post a table of all the results here once this is done.