Daetwyler Stephan, Mazloom-Farsibaf Hanieh, Zhou Felix Y, Segal Dagan, Sapoznik Etai, Chen Bingying, Westcott Jill M, Brekken Rolf A, Danuser Gaudenz, Fiolka Reto
Lyda Hill Department of Bioinformatics, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Nat Methods. 2025 Mar;22(3):569-578. doi: 10.1038/s41592-025-02598-2. Epub 2025 Feb 12.
Most biological processes, from development to pathogenesis, span multiple time and length scales. While light-sheet fluorescence microscopy has become a fast and efficient method for imaging organisms, cells and subcellular dynamics, simultaneous observations across all these scales have remained challenging. Moreover, continuous high-resolution imaging inside living organisms has mostly been limited to a few hours, as regions of interest quickly move out of view due to sample movement and growth. Here, we present a self-driving, multiresolution light-sheet microscope platform controlled by custom Python-based software, to simultaneously observe and quantify subcellular dynamics in the context of entire organisms in vitro and in vivo over hours of imaging. We apply the platform to the study of developmental processes, cancer invasion and metastasis, and we provide quantitative multiscale analysis of immune-cancer cell interactions in zebrafish xenografts.
大多数生物过程，从发育到发病机制，都跨越多个时间和长度尺度。虽然光片荧光显微镜已成为一种快速高效的生物体、细胞和亚细胞动力学成像方法，但在所有这些尺度上进行同步观察仍然具有挑战性。此外，由于样本移动和生长，感兴趣区域很快就会移出视野，因此在活生物体内进行连续高分辨率成像大多限于几个小时。在这里，我们展示了一个由基于Python的定制软件控制的自动驾驶多分辨率光片显微镜平台，用于在体外和体内对整个生物体进行数小时成像的背景下，同时观察和量化亚细胞动力学。我们将该平台应用于发育过程、癌症侵袭和转移的研究，并对斑马鱼异种移植中的免疫-癌细胞相互作用进行了定量多尺度分析。
