Three-dimensional imaging by deconvolution microscopy

Deconvolution is a computational method used to reduce out-of-focus fluores cence in three-dimensional (3D) microscope images. It can be applied in pri nciple to any type of microscope image but has most often been used to impr ove images from conventional fluorescence microscopes. Compared to other fo rms of 3D light microscopy, like confocal microscopy, the advantage of deco nvolution microscopy is that it can be accomplished at very low light level s, thus enabling multiple focal-plane imaging of light-sensitive living spe cimens over long time periods. Here we discuss the principles of deconvolut ion microscopy, describe different computational approaches for deconvoluti on, and discuss interpretation of deconvolved images with a particular emph asis on:what artifacts may arise. (C) 1999 Academic Press.