A comparison of image restoration approaches applied to three‐dimensional confocal and wide‐field fluorescence microscopy

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Abstract
We have compared different image restoration approaches for fluorescence microscopy. The most widely used algorithms were classified with a Bayesian theory according to the assumed noise model and the type of regularization imposed. We considered both Gaussian and Poisson models for the noise in combination with Tikhonov regularization, entropy regularization, Good's roughness and without regularization (maximum likelihood estimation). Simulations of fluorescence confocal imaging were used to examine the different noise models and regularization approaches using the mean squared error criterion. The assumption of a Gaussian noise model yielded only slightly higher errors than the Poisson model. Good's roughness was the best choice for the regularization. Furthermore, we compared simulated confocal and wide‐field data. In general, restored confocal data are superior to restored wide‐field data, but given sufficient higher signal level for the wide‐field data the restoration result may rival confocal data in quality. Finally, a visual comparison of experimental confocal and wide‐field data is presented.