Daetwyler Stephan, Chang Bo-Jui, Chen Bingying, Zhou Felix, Fiolka Reto
Lyda Hill Department for Bioinformatics, UT Southwestern Medical Center, 6000 Harry Hines BLVD, Dallas, TX 75390, USA.
bioRxiv. 2023 Aug 11:2023.08.10.552834. doi: 10.1101/2023.08.10.552834.
Understanding the intricate interplay and inter-connectivity of biological processes across an entire organism is important in various fields of biology, including cardiovascular research, neuroscience, and developmental biology. Here, we present a mesoscopic oblique plane microscope (OPM) that enables whole organism imaging with high speed and subcellular resolution. A microprism underneath the sample enhances the axial resolution and optical sectioning through total internal reflection of the light-sheet. Through rapid refocusing of the light-sheet, the imaging depth is extended up to threefold while keeping the axial resolution constant. Using low magnification objectives with a large field of view, we realize mesoscopic imaging over a volume of 3.7×1.5×1 mm with ~2.3 microns lateral and ~9.2 microns axial resolution. Applying the mesoscopic OPM, we demonstrate and whole organism imaging of the zebrafish vasculature and its endothelial nuclei, and blood flow dynamics at 12 Hz acquisition rate, resulting in a quantitative map of blood flow across the entire organism.
了解整个生物体中生物过程的复杂相互作用和相互联系在生物学的各个领域都很重要，包括心血管研究、神经科学和发育生物学。在这里，我们展示了一种介观斜平面显微镜（OPM），它能够以高速和亚细胞分辨率对整个生物体进行成像。样品下方的微棱镜通过光片的全内反射提高了轴向分辨率和光学切片能力。通过光片的快速重新聚焦，成像深度扩展了两倍，同时保持轴向分辨率不变。使用具有大视野的低倍物镜，我们实现了对体积为3.7×1.5×1毫米的区域进行介观成像，横向分辨率约为2.3微米，轴向分辨率约为9.2微米。应用介观OPM，我们展示了斑马鱼脉管系统及其内皮细胞核的整个生物体成像，以及在12 Hz采集速率下的血流动力学，从而生成了整个生物体的血流定量图。
