Kim Hyeonggeon, Natan Ryan, Chen Wei, Winans Amy M, Fan Jiang Lan, Isacoff Ehud, Ji Na
Opt Express. 2025 Jan 13;33(1):680-693. doi: 10.1364/OE.541033.
Adaptive optics (AO) improves the spatial resolution of microscopy by correcting optical aberrations. While its application has been well established in microscopy modalities utilizing a circular pupil, its adaptation to systems with non-circular pupils, such as Bessel-focus two-photon fluorescence microscopy (2PFM) with an annular pupil, remains relatively uncharted. Herein, we present a modal focal AO (MFAO) method for Bessel-focus 2PFM. Measuring and correcting aberration using a spatial light modulator placed in conjugation with the focal plane of the microscope objective, MFAO employs Zernike annular polynomials - a first in AO implementation - to achieve performance on par with a previous zonal AO method, but with a notably simplified optical configuration. We validated the performance of MFAO in correcting artificial and sample-induced aberrations, as well as in imaging of zebrafish larvae and mouse brains. By expanding the application of modal AO to annular pupils as well as aberration measurement and correction to a wavefront modulator at the objective focal plane, MFAO represents a notable advancement in the implementation of AO in microscopy.
自适应光学（AO）通过校正光学像差提高显微镜的空间分辨率。虽然其在利用圆形光瞳的显微镜模式中的应用已得到充分确立，但其在具有非圆形光瞳的系统中的应用，例如具有环形光瞳的贝塞尔聚焦双光子荧光显微镜（2PFM），仍相对未知。在此，我们提出一种用于贝塞尔聚焦2PFM的模态聚焦AO（MFAO）方法。MFAO使用与显微镜物镜焦平面共轭放置的空间光调制器来测量和校正像差，采用泽尼克环形多项式——这是AO实现中的首次应用——以实现与先前的区域AO方法相当的性能，但光学配置显著简化。我们验证了MFAO在校正人为和样品诱导的像差以及斑马鱼幼虫和小鼠大脑成像方面的性能。通过将模态AO的应用扩展到环形光瞳，并将像差测量和校正扩展到物镜焦平面处的波前调制器，MFAO代表了AO在显微镜应用中的显著进展。
