Davis Samuel, Sommernes Jon-Richard, Hambura Sebastian, Riedel Levin, Gil Alejandro, Ikmi Aissam, Ströhl Florian, Prevedel Robert
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Department of Physics and Technology, UiT The Arctic University of Norway, Tromsø, Norway.
Biomed Opt Express. 2024 Nov 6;15(12):6715-6724. doi: 10.1364/BOE.537262. eCollection 2024 Dec 1.
Rapid three-dimensional imaging over extended fields of view (FOVs) is crucial to the study of organism-wide systems and biological processes . Selective-plane illumination microscopy (SPIM) is a powerful method for high spatio-temporal resolution imaging of such biological specimens. However, typical SPIM implementations preclude conventional sample mounting and have anisotropic imaging performance, in particular when designed for large FOVs over 1 mm diameter. Here, we introduce axial sweeping of the illumination into a non-orthogonal dual-objective oblique plane microscope (OPM) design, thereby enabling the observation of freely moving animals over millimeter-sized FOVs, at close to isotropic, sub-cellular resolution. We apply our mesoscopic axially swept OPM (MASOPM) to image the behavioral dynamics of the sea anemone over 1 × 0.7 × 0.4 mm at 1.7 × 2.6 × 3.7 µm resolution and 0.5 Hz volume rate.
在扩展视野（FOV）上进行快速三维成像对于研究整个生物体系统和生物过程至关重要。选择性平面照明显微镜（SPIM）是一种用于对这类生物样本进行高时空分辨率成像的强大方法。然而，典型的SPIM实现方式排除了传统的样本安装方式，并且具有各向异性的成像性能，特别是当设计用于直径超过1毫米的大视野时。在这里，我们将照明的轴向扫描引入非正交双物镜斜平面显微镜（OPM）设计中，从而能够以接近各向同性的亚细胞分辨率观察毫米级视野内自由移动的动物。我们应用我们的介观轴向扫描OPM（MASOPM）以1.7×2.6×3.7微米的分辨率和0.5赫兹的体积速率对海葵在1×0.7×0.4毫米范围内的行为动态进行成像。
