Imaging of cellular dynamicsin vitroandin situ: from a whole organism to sub-cellular imaging with self-driving, multi-scale microscopy

Stephan Daetwyler, Hanieh Mazloom-Farsibaf, Felix Y. Zhou, Dagan Segal, Etai Sapoznik, Jill M. Westcott, Rolf A. Brekken, Gaudenz Danuser, Reto Fiolka
2024 · doi:10.1101/2024.02.28.582579
preprint OA: gold CC-BY-NC-ND-4.0
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Abstract

Many biological processes span multiple time and length scales, including developmental processes and cancer metastasis. While light-sheet fluorescence microscopy (LSFM) has become a fast and efficient method for imaging of organisms, cells and sub-cellular dynamics, simultaneous observations across these scales have remained challenging. Moreover, continuous high-resolution imaging inside living organisms has mostly been limited to few hours as regions of interest quickly move out of view due to sample movement and growth. Here, we present a self-driving, multi-resolution light-sheet microscope platform controlled by a custom Python-based software, to simultaneous observe and quantify sub-cellular dynamics and 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 multi-scale analysis of immune-cancer cell interactions in zebrafish xenografts.

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License: CC-BY-NC-ND-4.0