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Super-resolved live-cell imaging using Random Illumination Microscopy

Simon Labouesse
  • Function : Author
Marc Allain
Emmanuel Martin
Renaud Poincloux
Anaïs Bouissou
  • Function : Author
Sylvain Cantaloube
  • Function : Author
Elise Courtaux
  • Function : Author
Elodie Vega
  • Function : Author
Tong Li
  • Function : Author
Aude Guénolé
  • Function : Author
Christian Rouvière
  • Function : Author
Sophie Allard
  • Function : Author
Nathalie Campo
  • Function : Author
Xiaobo Wang
  • Function : Author
Grégoire Michaux
Mathieu Pinot
Roland Le Borgne
  • Function : Author
Sylvie Tournier
Jérôme Idier
Anne Sentenac
  • Function : Author

Abstract

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 .

Dates and versions

hal-03101965 , version 1 (07-01-2021)

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Thomas Mangeat, Simon Labouesse, Marc Allain, Emmanuel Martin, Renaud Poincloux, et al.. Super-resolved live-cell imaging using Random Illumination Microscopy. 2021. ⟨hal-03101965⟩
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