Two-photon microscopy through scattering media harnessing speckle autocorrelation

Abstract

Two-photon (2P) microscopy is a powerful technique for deep-tissue fluorescence imaging; however, tissue scattering limits its effectiveness for depth imaging using conventional approaches. Despite typical strategies having been put forward to extend depth imaging capabilities based on wave-front shaping (WFS), computationally recovering images remains a significant challenge using 2P signal. In this work, we demonstrate the successful reconstruction of fluorescent objects behind scattering layers using 2P microscopy, utilizing the optical memory effect (ME) along with the speckle autocorrelation technique and a phase retrieval algorithm. Our results highlight the effectiveness of this method, offering significant potential for improving depth imaging capabilities in 2P microscopy through scattering media.