Re: General question: Software vs. hardware

Posted by Talley Lambert on
URL: http://confocal-microscopy-list.275.s1.nabble.com/General-question-Software-vs-hardware-tp7589316p7589326.html

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As Mike said, this is indeed a very old debate, but there are well-characterized, objective costs and benefits to each approach (scientific, not just monetary).  Deconvolution is not just "computational confocal at the expense of artifacts"... it's a false comparison.  By rejecting out-of-focus fluorescence, confocal microscopes reduce the *shot noise* contributed to the image by background.  Deconvolution, by contrast attempts "reassign" that out of focus information (provided you have a very accurate representation of the actual PSF in your sample...), but there will come a point with thicker samples at which the shot noise contributed by out-of-focus fluorescence overwhelms the SNR in the image, and deconvolution will fail (figure 4 in the first paper below).  However, for thin samples with minimal out-of-focus fluorescence, the increased collection efficiency and minimized illumination/detector noise of widefield+decon has benefits for detection of weak signals (figure 2 in the paper below).

This tradeoff was well-characterized by Swedlow and Murray
https://www.ncbi.nlm.nih.gov/pubmed/11830634

...and followed up with a treatment on the photon-efficiency of different optical sectioning techniques:
 https://www.ncbi.nlm.nih.gov/pubmed/18045334

As usual, there is no one technique that is universally "better" or preferable.  It will depend on the samples you are imaging and the relative levels of in-focus and out-of-focus information.