Digital light microscopy: prerequisite for optimum contrast enhancement and increase of resolution

Since the reproducible production of microscope objectives was enabled by the lens calculations of Ernst Abbe in 1872, various attempts have been made to further increase the resolution of light microscopes. Apart from the improvements on the optical side, especially the introduction of fluorescence methods, the use of digital cameras connected to computers have brought us close to the theoretical limits of optical resolution. Due to improved speed and memory capacity of modern computers mathematical methods can be applied to stored three-dimensional (3D) sequences of digital images which, in addition to just contrast and edge enhancement, may result in the case of real 3D deconvolution, even in the visualisation of structures beyond the theoretical limitations of light optical resolution.