A Workingperson’s Guide to Deconvolution in Light Microscopy

TLDR: The imaging process in the fluorescence microscope is reviewed and then how the various deconvolution methods work are discussed, to discuss imaging artifacts and how to minimize them.

Abstract: The fluorescence microscope is routinel y used to study cellular structure in many bi o medical research laboratories and is i n creasingly used as a quantitative assay sy s tem for cellular dynamics. One of the majo r causes of image degradation in the fluore s cence microscope is blurring. Deconvol u tion algorithms use a model of the micr o scope imaging process to either subtract o r reassign out-of-focus blur. A variety of a l gorithms are now commercially available , each with its own characteristic advantage s and disadvantages. In this article, we r e view the imaging process in the fluore s cence microscope and then discuss how th e various deconvolution methods work. Fina l ly, we provide a summary of practical tip s for using deconvolution and discuss ima g ing artifacts and how to minimize them .