Replica-Assisted Super-Resolution Fluorescence Imaging in Scattering Media

Abstract

Far-field super-resolution fluorescence microscopy has been rapidly developed for applications ranging from cell biology to nanomaterials. However, it remains a significant challenge to achieve super-resolution imaging at depth in opaque materials. In this study, we present a super-resolution microscopy technique for imaging hidden fluorescent objects through scattering media, started by exploiting the inherent object replica generation arising from the memory effect, i.e., the seemingly informationless emission speckle can be regarded as a random superposition of multiple object copies. Inspired by the concept of super-resolution optical fluctuation imaging, we use temporally fluctuating speckles to excite fluorescence signals and perform high-order cumulant analysis on the fluctuation, which can not only improve the image resolution but also increase the speckle contrast to isolate only the bright object replicas. A super-resolved image can be finally retrieved by simply unmixing the sparsely distributed replicas with their location map. This methodology allows one to overcome scattering and achieve robust super-resolution fluorescence imaging, circumventing the need for heavy computational steps.