3D quantitative imaging at different scales using single objective light-sheet microscopy

Assessing protein organization and dynamics in their native cellular context provides invaluable insights into their activities and functions. Fluorescence microscopy has drastically improved and diversified over the last 20 years, allowing major discoveries in cell biology, neuroscience and developmental biology. It is today possible to monitor the evolution of fluorescent markers over a period of time ranging between milliseconds up to several days with down to single molecule spatial resolution, in very diverse living biological systems ranging from single cells up to whole embryos. However, depending on the biological question, one usually requires to use several, expensive and often very sophisticated imaging techniques.

I will present the capabilities and requirements of the soSPIM (single-objective Selective Plane Illumination Microscopy) imaging technique to: i) probe the 3D nanoscale organization of proteins in depth at the single-molecule level, and ii) achieve quantitative high-content imaging of living and fixed 3D cell-cultures (spheroids/organoids) with unprecedented throughput. soSPIM is a light-sheet microscopy technique operating on a standard mono-objective inverted microscope. It relies on dedicated microchips embedding arrays of microwells flanked with 45° micro-mirrors acting as light guides and culture vessels.

[1] Galland et al., 3D high- and super-resolution imaging using single-objective SPIM, Nat Methods, 12 (2015) 641-644.
[2] Singh et al., 3D Protein Dynamics in the Cell Nucleus. Biophys J. 2017.
[3] Beghin et al., Automated high-speed 3D imaging of organoid cultures with multi-scale phenotypic quantification. Nat Methods. 2022.