# I need to deconvolve the image of a fluorescent bead without the Point Spread Function in MATLAB

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.

Alternative Solution

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.

• Crossposted to physics.stackexchange.com/q/232098/2451 and dsp.stackexchange.com/q/28510 Jan 27 '16 at 15:55
• @Qmechanic crossposted to SO at stackoverflow.com/questions/35041093
– SDG
Jan 27 '16 at 15:57
• Your problem goes under the name "blind deconvolution". It's a pretty hard problem but there are methods out there that you can try.
– Dirk
Jan 27 '16 at 20:47
• @Dirk I have already tried blind deconvolution, but it does not give the whole picture about the best possible deconvolution method with the microscope I am working with. I have added a bit to the question, if you would like to see.
– SDG
Jan 28 '16 at 4:42