Jean-Baptiste Sibarita, Institute for Interdisciplinary Neuroscience, Bordeaux Neurocampus, CNRS, France
Deciphering protein organization and dynamics at the nanoscale using multidimensional single molecule localization 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 verydiverse living biological systems ranging from isolated cells, up to whole organisms. Single molecule localization microscopy (SMLM) is a powerful super-resolution microscopy technique allowing monitoring the nanoscale co-organization and dynamics of fluorescent proteins in their native environment. However, it remains limited in its throughput, its capability to investigate 3D complex biological samples, and the quantifications that can be gathered.I will present our recent achievements on quantitative 3D localization using the soSPIM (single-objective Selective Plane Illumination Microscopy) imaging technique and the Point-Cloud Analyst, to probe the 3D nanoscale organization of proteins in depth in isolated cells and organotypic 3D cell cultures. 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. Galland et al., 3D high-and super-resolution imaging using single-objective SPIM, Nat Methods, 12 (2015). Levet et al., SR-Tesseler: a method to segment and quantify localization-based super-resolution microscopy data. Nat Methods. 2015. Beghin et al.,Automated high-speed 3D imaging of organoid cultures with multi-scale phenotypic quantification. Nat Methods. 2022. SIBARITA_JB_Abstract_…e_BrainC
JB Sibarita has a PhD thesis in Physics and is expert in live cell microscopy and image analysis. He has co-headed and developed the “Cellular and Tissular Imaging Platform” of the Institut Curie, Paris, for 12 years. In 2009, he created his CNRS R&D team “Quantitative Imaging of the Cell” in the newly created Interdisciplinary Institute of Neuroscience, Bordeaux. Together with the physicists, computer scientists and biologists within the team, they aim at developing novel quantitative imaging techniques to decipher the living cell activity at the nanoscale level, in a high throughput context. In 2018, he spent a year at the National University of Singapore to co-develop the single objective light-sheet microscope in the context of 3D cell culture.
During the last 15 years, JB Sibarita has initiated several academic and industrial partnerships, based on the developments achieved in his lab. He is at the origin of 5 industrial technology transfers as single or main author and 3 patents. He was awarded with the CNRS crystal medal in 2006 and a chaire d’excellence of the Regional Council of Aquitaine in 2009.