A variety of modern super-resolution microscopy methods provide researchers with previously inconceivable biological sample imaging opportunities at a molecular resolution. All of these techniques excel at imaging samples that are close to the coverslip, however imaging at large depths remains a challenge due to aberrations caused by the sample, diminishing the resolution of the microscope. Originating in astro-imaging, the adaptive optics (AO) approach for wavefront shaping using a deformable mirror is gaining momentum in modern microscopy as a convenient approach for wavefront control. AO has the ability not only to correct aberrations but also enables engineering of the PSF shape, allowing localization of the emitter axial position over several microns. In this study, we demonstrate remote focusing as another AO benefit for super-resolution microscopy. We show the ability to record volumetric data (45 × 45 × 10 µm), while keeping the sample axially stabilized using a standard widefield setup with an adaptive optics addon. We processed the data with single-molecule localization routines and/or computed spatiotemporal correlations, demonstrating subdiffraction resolution.

中文翻译：

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自适应光学通过远程聚焦实现多模式 3D 超分辨率显微镜

各种现代超分辨率显微镜方法为研究人员提供了以前无法想象的分子分辨率的生物样品成像机会。所有这些技术都擅长对靠近盖玻片的样品进行成像，但是由于样品引起的像差，大深度成像仍然是一个挑战，降低了显微镜的分辨率。起源于天文成像，使用可变形镜进行波前整形的自适应光学 (AO) 方法在现代显微镜中作为一种方便的波前控制方法正在获得动力。AO 不仅能够校正像差，还能够设计 PSF 形状，从而可以将发射器轴向位置定位在几微米内。在这项研究中，我们展示了远程聚焦是超分辨率显微镜的另一个 AO 优势。我们展示了记录体积数据 (45 × 45 × 10 µm) 的能力，同时使用带有自适应光学插件的标准宽场设置保持样品轴向稳定。我们使用单分子定位程序和/或计算的时空相关性处理数据，展示了亚衍射分辨率。