3D visualization in multifocus fluorescence microscopy

Limited depth-of-field can be overcome through computational optical imaging. In this work, a custom built fluorescence microscope with a focus electrically tunable lens is used for the acquisition of a multifocus image sequence (z-stack) of a 3D fluorescent sample. Image registration between the acquired images is often needed as a preprocessing step before the reconstruction of images with new characteristics. Then a multifocus image fusion algorithm is implemented to accomplish all-in-focus image reconstruction from the registered z-stack. Also computational perspective shifts that allow to the reconstruction of stereoscopic pairs of the sample and its three-dimensional visualization are implemented.