Deconvolution Software for Modern Fluorescence Microscopy

This software invention pertains to Joint Richardson-Lucy (RL) deconvolution methods used to combine multiple images of an object into a single image for improving resolution in modern fluorescence microscopy. RL deconvolution merges images with very different point spread functions, such as in multi-view light-sheet microscopes, while preserving the best resolution information present in each image. RL deconvolution is also easily applied to merge high-resolution, high noise images with low-resolution, low noise images, relevant when complementing conventional microscopy with localization microscopy. The technique can be performed on images produced via different simulated illumination patterns, relevant to structured illumination microscopy (SIM) and image scanning microscopy (ISM) resulting in image qualities at least as good as standard inversion algorithms, but follows a simpler protocol that requires little mathematical insight. RL deconvolution can also be used to merge a series of several images with varying signal and resolution levels. This combination is relevant to gated stimulated-emission depletion (STED) microscopy and shows that high-quality image merges are possible even in cases where no explicit inversion algorithm is known.