Entropy-Based Super-Resolution Imaging (ESI): From Disorder to Fine Detail

We introduce a novel and universal method for fast optical high, as well as super-resolution imaging. Our method is based on reconstructing super-resolved images from conventional image sequences containing rapid random signal fluctuations. Such sequences could be obtained from either wide-field single-molecule blinking experiments or rapid image sequences with fluorophores undergoing random intensity fluctuations. By calculating the local entropy (H) and cross-entropy (xH) values pixel-by-pixel, weighted with higher order statistics (HOS), a new image with pixel intensities representing the true information content in the time series is obtained. We show that analyzing image sequences by this formalism enables the reconstruction of super-resolved images, where the optical resolution that can be achieved depends only on the number of input frames and the higher order moments used for the calculation. We find that the acquisition of <100 frames per sequence is sufficient to reconstruct super-resolved images of entire cells. We also demonstrate that not only on off switching of the fluorescent. dyes, but also other dynamic events, that is, photobleaching, can be exploited for efficient and high-resolution image reconstructions. This method opens up the potential to obtain super-resolved images from most wide-field fluorescence microscopy systems. By providing a universal Fiji-plugin most users of high-end fluorescence microscopy systems will now benefit from this easy-to-use super-resolution optical microscopy method.