Compressive Fourier-Domain Intensity Coupling (C-FOCUS) enables near-millimeter deep imaging in the intact mouse brain in vivo
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Two-photon microscopy is a powerful tool for in vivo imaging, but its imaging depth is typically limited to a few hundred microns due to tissue scattering, even with existing scattering correction techniques. Moreover, most active scattering correction methods are restricted to small regions by the optical memory effect. Here, we introduce compressive Fourier-domain intensity coupling for scattering correction (C-FOCUS), an active scattering correction approach that integrates Fourier-domain intensity modulation with compressive sensing for two-photon microscopy. Using C-FOCUS, we demonstrate high-resolution imaging of YFP labeled neurons and FITC-labeled blood vessels at depths exceeding 900 µm in the intact mouse brain in vivo. Furthermore, we achieve transcranial imaging of YFP-labeled dendritic structures through the intact adult mouse skull. C-FOCUS enables high-contrast fluorescence imaging at depths previously inaccessible using two-photon microscopy with 1035 nm excitation, enhancing fluorescence intensity by over 20-fold compared to uncorrected imaging. C-FOCUS provides a broadly applicable strategy for rapid, deep-tissue optical imaging in vivo.