Superresolution Microscopy, Institute of Molecular Biology (IMB), D-55128, Mainz, Germany.
Physics Department University Mainz (JGU), D-55128, Mainz, Germany.
Sci Rep. 2017 Jun 16;7(1):3685. doi: 10.1038/s41598-017-03743-4.
The limits of conventional light microscopy ("Abbe-Limit") depend critically on the numerical aperture (NA) of the objective lens. Imaging at large working distances or a large field-of-view typically requires low NA objectives, thereby reducing the optical resolution to the multi micrometer range. Based on numerical simulations of the intensity field distribution, we present an illumination concept for a super-resolution microscope which allows a three dimensional (3D) optical resolution around 150 nm for working distances up to the centimeter regime. In principle, the system allows great flexibility, because the illumination concept can be used to approximate the point-spread-function of conventional microscope optics, with the additional benefit of a customizable pupil function. Compared with the Abbe-limit using an objective lens with such a large working distance, a volume resolution enhancement potential in the order of 10 is estimated.
传统光学显微镜的极限（“阿贝极限”）主要取决于物镜的数值孔径（NA）。在大工作距离或大视场进行成像通常需要低数值孔径的物镜，从而将光学分辨率降低到多微米的范围。基于强度场分布的数值模拟，我们提出了一种用于超分辨率显微镜的照明概念，该概念允许在高达厘米范围的工作距离下实现三维（3D）光学分辨率约为 150nm。原则上，该系统具有很大的灵活性，因为照明概念可用于近似传统显微镜光学器件的点扩散函数，并且还具有可定制的光瞳函数的额外优势。与使用具有如此大工作距离的物镜的阿贝极限相比，预计体积分辨率的提高潜力约为 10 倍。
