High-resolution scanning fluorescence imaging through scattering via speckle replica alignment and variance computation

Abstract

Fluorescence imaging is an essential diagnostic tool in many fields, but diffraction-limited optical imaging at depth is limited by scattering. Here, we present a method based on multiple random illuminations, combined with a computational framework that retrieves high-resolution images by aligning local speckle replicas and computing their pixel-wise variance. We demonstrate its versatility in two regimes: linear wide-field one-photon (1P) fluorescence imaging and nonlinear two-photon (2P) fluorescence imaging where the object is excited by a scanned speckle field and detected with a single-pixel detector. This approach outperforms standard autocorrelation techniques in terms of resolution and convergence.