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Super-resolved live-cell imaging using Random Illumination Microscopy

Thomas Mangeat, Simon Labouesse, Marc Allain, Emmanuel Martin, Renaud Poincloux, Anaïs Bouissou, Sylvain Cantaloube, Elise Courtaux, Elodie Vega, Tong Li, Aude Guénolé, Christian Rouvière, Sophie Allard, Nathalie Campo, Magali Suzanne, Xiaobo Wang, View ORCID ProfileGrégoire Michaux, Mathieu Pinot, Roland Le Borgne, Sylvie Tournier, Jérôme Idier, Anne Sentenac
doi: https://doi.org/10.1101/2020.01.27.905083
Thomas Mangeat
LITC Core Facility, Centre de Biologie Integrative, Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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  • For correspondence: thomas.mangeat@univ-tlse3.fr
Simon Labouesse
Institut Fresnel, Aix Marseille Univ, CNRS, Centrale Marseille, Marseille, France
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Marc Allain
Institut Fresnel, Aix Marseille Univ, CNRS, Centrale Marseille, Marseille, France
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Emmanuel Martin
LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Renaud Poincloux
Institut de Pharmacologie et de Biologie Structurale (IPBS), Université de Toulouse, CNRS, UPS, Toulouse, France
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Anaïs Bouissou
Institut de Pharmacologie et de Biologie Structurale (IPBS), Université de Toulouse, CNRS, UPS, Toulouse, France
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Sylvain Cantaloube
LITC Core Facility, Centre de Biologie Integrative, Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Elise Courtaux
Laboratoire de Microbiologie et Génétique Moléculaires (LMGM), UMR5100, Centre de Biologie Intégrative (CBI), CNRS, Toulouse, France
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Elodie Vega
Institut de Pharmacologie et de Biologie Structurale (IPBS), Université de Toulouse, CNRS, UPS, Toulouse, France
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Tong Li
LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Aude Guénolé
LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Christian Rouvière
LITC Core Facility, Centre de Biologie Integrative, Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Sophie Allard
INSERM Université de Toulouse, UPS, CNRS, Centre de Physiopathologie de Toulouse Purpan (CPTP), Toulouse, France
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Nathalie Campo
Laboratoire de Microbiologie et Génétique Moléculaires (LMGM), UMR5100, Centre de Biologie Intégrative (CBI), CNRS, Toulouse, France
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Magali Suzanne
LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Xiaobo Wang
LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Grégoire Michaux
Univ Rennes, CNRS, Institut de Génétique et Développement de Rennes (IGDR) - UMR 6290, 35000 Rennes, France
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  • ORCID record for Grégoire Michaux
Mathieu Pinot
Univ Rennes, CNRS, Institut de Génétique et Développement de Rennes (IGDR) - UMR 6290, 35000 Rennes, France
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Roland Le Borgne
Univ Rennes, CNRS, Institut de Génétique et Développement de Rennes (IGDR) - UMR 6290, 35000 Rennes, France
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Sylvie Tournier
LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, 31062 France
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Jérôme Idier
LS2N, CNRS UMR 6004, 1 rue de la Noë, F44321 Nantes Cedex 3, France
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Anne Sentenac
Institut Fresnel, Aix Marseille Univ, CNRS, Centrale Marseille, Marseille, France
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Summary

Super-resolution fluorescence microscopy has been instrumental to progress in biology. Yet, the photo-induced toxicity, the loss of resolution into scattering samples or the complexity of the experimental setups curtail its general use for functional cell imaging. Here, we describe a new technology for tissue imaging reaching a 114nm/8Hz resolution at 30 µm depth. Random Illumination Microscopy (RIM) consists in shining the sample with uncontrolled speckles and extracting a high-fidelity super-resolved image from the variance of the data using a reconstruction scheme accounting for the spatial correlation of the illuminations. Super-resolution unaffected by optical aberrations, undetectable phototoxicity, fast image acquisition rate and ease of use, altogether, make RIM ideally suited for functional live cell imaging in situ. RIM ability to image molecular and cellular processes in three dimensions and at high resolution is demonstrated in a wide range of biological situations such as the motion of Myosin II minifilaments in Drosophila.

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Posted January 27, 2020.
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Super-resolved live-cell imaging using Random Illumination Microscopy
Thomas Mangeat, Simon Labouesse, Marc Allain, Emmanuel Martin, Renaud Poincloux, Anaïs Bouissou, Sylvain Cantaloube, Elise Courtaux, Elodie Vega, Tong Li, Aude Guénolé, Christian Rouvière, Sophie Allard, Nathalie Campo, Magali Suzanne, Xiaobo Wang, Grégoire Michaux, Mathieu Pinot, Roland Le Borgne, Sylvie Tournier, Jérôme Idier, Anne Sentenac
bioRxiv 2020.01.27.905083; doi: https://doi.org/10.1101/2020.01.27.905083
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Super-resolved live-cell imaging using Random Illumination Microscopy
Thomas Mangeat, Simon Labouesse, Marc Allain, Emmanuel Martin, Renaud Poincloux, Anaïs Bouissou, Sylvain Cantaloube, Elise Courtaux, Elodie Vega, Tong Li, Aude Guénolé, Christian Rouvière, Sophie Allard, Nathalie Campo, Magali Suzanne, Xiaobo Wang, Grégoire Michaux, Mathieu Pinot, Roland Le Borgne, Sylvie Tournier, Jérôme Idier, Anne Sentenac
bioRxiv 2020.01.27.905083; doi: https://doi.org/10.1101/2020.01.27.905083

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