Mcgorty Ryan, Xie Dan, Huang Bo
Opt Express. 2017 Jul 24;25(15):17798-17810. doi: 10.1364/OE.25.017798.
Selective-plane illumination microscopy (SPIM) provides unparalleled advantages for the volumetric imaging of living organisms over extended times. However, the spatial configuration of a SPIM system often limits its compatibility with many widely used biological sample holders such as multi-well chambers and plates. To solve this problem, we developed a high numerical aperture (NA) open-top configuration that places both the excitation and detection objectives on the opposite of the sample coverglass. We carried out a theoretical calculation to analyze the structure of the system-induced aberrations. We then experimentally compensated the system aberrations using adaptive optics combined with static optical components, demonstrating near-diffraction-limited performance in imaging fluorescently labeled cells.
选择性平面照明显微镜（SPIM）在长时间对活生物体进行体积成像方面具有无与伦比的优势。然而，SPIM系统的空间配置常常限制了它与许多广泛使用的生物样品支架（如多孔腔室和培养皿）的兼容性。为了解决这个问题，我们开发了一种高数值孔径（NA）的开放式顶部配置，将激发物镜和检测物镜都放置在样品盖玻片的相对侧。我们进行了理论计算以分析系统诱导像差的结构。然后，我们通过结合自适应光学和静态光学元件对系统像差进行了实验补偿，在对荧光标记细胞成像时展示了近衍射极限的性能。
