Precise 3D computer-generated holography based on non-convex optimization with spherical aberration compensation (SAC-NOVO) for two-photon optogenetics

Computer-generated holography (CGH) has been adopted in two-photon optogenetics as a promising technique for selective excitation of neural ensembles. However, 3D CGH by nonconvex optimization, the state of art method, is susceptible to imprecise axial positioning, due to the quadratic phase approximation in 3D target generation. Even though the misplacement of targets in conventional CGH can be solved by pre-calibration, it still suffers from low efficiency and poor axial resolution of two-photon excitation. Here, we propose a novel CGH method based on non-convex optimization with spherical aberration compensation (SAC-NOVO). Through numerical simulations and two-photon excitation experiments, we verify that SAC-NOVO could achieve precise axial positioning for single and multiple expanded disk patterns, while ensuring high two-photon excitation efficiency. Besides, we experimentally show that SAC-NOVO enables the suppression of dark target areas. This work shows the superiority of SAC-NOVO for two-photon optogenetics.