3D live imaging at different scales using single-objective SPIM

Session

LST.10 - Lightsheet illumination/detection strategies to yield higher speed, higher resolution and higher throughput in Bioimaging

Authors

Hisham Forriere (2), Anne Beghin (1), Marine Cabillic (2), Corey Butler (2), Gianluca Grenci (1), Rémi Galland (2), Virgile Viasnoff (1), Jean-Baptiste Sibarita (2)

Affiliations

1. National University of Singapore
2. University of Bordeaux

Keywords

Light-Sheet Microscopy, Single Molecule Localization Microscopy, Organoid, Live Imaging

Abstract text

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 require to use several, expensive and often very sophisticated imaging techniques.

We will present the capabilities and requirements of the soSPIM (single-objective Selective Plane Illumination Microscopy) imaging technique [1] to: i) probe the 3D nanoscale organization of proteins in depth at the single-molecule level, and ii) achieve high-content screening of living spheroids/organoid with unprecedented throughput. soSPIM is a light-sheet microscopy technique operating on a standard mono objective inverted microscope thanks to the use of a dedicated microchips embedding arrays of microwells flanked with 45° micro-mirrors.

References

[1] R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, J.B. Sibarita, 3D high- and super-resolution imaging using single-objective SPIM, Nat Methods, 12 (2015) 641-644.