Volumetric light-sheet data of freely moving sea anemone

Several important questions in biology require non-invasive and three-dimensional imaging techniques with appropriate spatiotemporal resolution that permit live organisms to move in an unconstrained fashion over an extended field-of-view. While selective-plane illumination microscopy (SPIM) has emerged as a powerful method to observe live biological specimens at high spatio-temporal resolution, typical implementations often necessitate constraining sample mounting or lack the required volumetric speed. Here, we report on an open-top, dual-objective oblique plane microscope (OPM) capable of observing millimeter sized, freely moving animals at cellular resolution. We demonstrate the capabilities of our mesoscopic OPM (MesOPM) by imaging the behavioural dynamics of the sea anemone Nematostella vectensis over 1.56 × 1.56 × 0.25 mm at 1.5 × 2.8 × 5.3µm resolution and 0.5Hz volume rate.

生物学领域诸多重要研究问题，均要求采用具备适配时空分辨率的无创三维成像技术，以支持活体生物在开阔视场内无约束地自由活动。尽管选择性平面照明显微镜（selective-plane illumination microscopy, SPIM）已成为实现高时空分辨率活体生物标本观测的有力手段，但典型系统配置往往需要对样本进行约束式固定，或无法满足所需的体成像速度。本研究报道了一款开放式双物镜斜平面显微镜（oblique plane microscope, OPM），其可在细胞分辨率下对毫米级大小的自由活动动物开展成像。我们通过对星状海葵（Nematostella vectensis）的行为动态进行成像，验证了所研发的介观斜平面显微镜（mesoscopic OPM, MesOPM）的性能：成像视场达1.56 × 1.56 × 0.25 mm，空间分辨率为1.5 × 2.8 × 5.3 µm，体成像帧率达0.5Hz。