Posted by
John Oreopoulos on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Deconvolution-of-Confocal-Images-was-Airy-Units-tp6946310p6946351.html
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Peter, there are some advantages to deconvolving even confocal imaging data. Until I took Pawley's course, I too had always seen these two imaging techniques as mutually exclusive, but that's not the case.
Here is a passage from Chapter 2 of Pawley's Hanbook:
"Although deconvolution and confocal imaging are often seen as competing methods aimed at the same goal, namely producing images of 3D stain distributions, in fact, they are not exclusive, and there is much to be said for combining them. No only does deconvolution suppress the "single-pixel" features created by Poisson noise, it also effectively averages the signal in the Nyquist-sampled image of a point object. In other words, it has the same effect of reducing the uncertainty in the estimate of the brightness in individual voxels as Kalman averaging for 64 to 125 frames. This point is so important that the present edition of this volume devotes an entire chapter to it: Chapter 25."
Now, having repeated that passage, can I say that I deconvolve my confocal data all the time? No, mainly due to lack of access to deconvolution software, and partly due to time constraints usually. But I can assure you that your images will look a bit better after deconvolving them. Apparently all the best confocalists do this, but I seldom see it practiced in the literature.
Have a look at Chapter 25 of his book. It's quite interesting.
John Oreopoulos
Research Assistant
Spectral Applied Research
Richmond Hill, Ontario
Canada
www.spectral.ca
On 2011-10-30, at 4:09 PM, Peter Werner wrote:
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> To join, leave or search the confocal microscopy listserv, go to:
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> An interesting point was made here by Jim Pawley:
>
>> I agree that sampling a bit higher than Nyquist never hurts, especially if you deconvolve (as you always should), but I think that it is a mistake to think that one can "separate" out the noise by decon. I think that noise is pretty fundamental.
>
> I had always heard that if you're doing confocal microscopy, at least point-scanning confocal with a pinhole size of 1AU or smaller, that deconvolution was superfluous, because you shouldn't be getting out of focus light. So what is gained by deconvolution when one is sampling voxel by voxel?
>
> Peter G. Werner
> Merritt College Microscopy Program