Second-harmonic generation microscopy with synthetic aperture and computational adaptive optics

Second -harmonic generation (SHG) microscopy is a powerful label -free imaging tool widely used to visualize collagen and muscle in biological tissues. However, traditional laser -scanning SHG microscopy requiring voxel scanning is time -intensive. Wide -field SHG microscopy was designed to bypass this restriction, but its application to deep tissue imaging is limited due to vulnerability to scattering and sample -induced aberrations. We introduce synthetic aperture SHG (SA-SHG) microscopy to attenuate the effect of multiple scattering noises. Our SA-SHG method coherently integrates amplitude and phase maps of wide -field SHG fields taken for different illumination angles, thereby enhancing the signal-to-noise ratio. We also develop computational adaptive optics SHG (CAO-SHG) microscopy to computationally correct the sample -induced aberrations. Our algorithm optimizes SHG fields' aperture synthesis to identify aberration maps, enabling the restoration of diffraction -limited imaging. We successfully apply this approach to real biological samples, demonstrating its potential for high -resolution imaging in complex biological environments. (c) 2024 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement