Wide-Field Multiphoton Imaging Through Scattering Media Without Correction

TL;DR

This paper introduces a novel wide-field two-photon imaging method that penetrates scattering media without prior correction, using temporal focusing and single-pixel detection to achieve deeper imaging with better signal quality.

Contribution

The authors present an innovative imaging scheme that does not require prior knowledge of the medium, enabling deeper imaging through scattering media with improved signal-to-background ratio.

Findings

Achieves imaging up to seven scattering mean free paths deep.

Provides up to fivefold improvement in signal-to-background ratio.

Reduces photobleaching compared to standard methods.

Abstract

Optical approaches to fluorescent, spectroscopic, and morphological imaging have made exceptional advances in the last decade. Super-resolution imaging and wide-field multiphoton imaging are now underpinning major advances across the biomedical sciences. While the advances have been startling, the key unmet challenge to date in all forms of optical imaging is to penetrate deeper. A number of schemes implement aberration correction or the use of complex photonics to address this need. In contrast, we approach this challenge by implementing a scheme that requires no a priori information about the medium nor its properties. Exploiting temporal focusing and single-pixel detection in our innovative scheme, we obtain wide-field two-photon images through various turbid media including a scattering phantom and tissue reaching a depth of up to seven scattering mean free path lengths. Our results show that it competes favorably with standard point-scanning two-photon imaging, with up to a fivefold improvement in signal-to-background ratio while showing significantly lower photobleaching.