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

TL;DR

This paper introduces a computational method for high-resolution fluorescence imaging through scattering media by aligning speckle replicas and analyzing variance, applicable to both 1P and 2P fluorescence imaging.

Contribution

The authors develop a novel speckle replica alignment and variance computation framework that enhances resolution in scattering fluorescence imaging, surpassing traditional autocorrelation methods.

Findings

Outperforms standard autocorrelation in resolution and convergence.

Effective in both linear 1P and nonlinear 2P fluorescence imaging regimes.

Demonstrates versatility and improved imaging quality through scattering media.

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.