There has been much provocative discussion over the past few years about the advantages and limitations of deconvolution methods. In this tutorial we will provide a reasonable understanding of the underlying theoretical principles of deconvolution, with an aim at understanding expected advantages and limitations. These principles will be presented for a mixed audience composed of biologists, neuroscientists, engineers, physicists and other scientists and graduate students. These principles will be presented in the context of both widefield and confocal microscopy deconvolution.

The basic principles of Fourier optics will be presented. From these principles, we can explain why it is that the confocal microscope provides a superior in-plane and axial (z-axis) resolving power, compared to the widefield microscope. In particular, for example, the Fourier transform of a 3D widefield image contains a missing cone region (to be explained in the tutorial), and this missing cone region effectively spoils the axial resolution of unprocessed widefield data.