Super-resolved live-cell imaging using Random Illumination Microscopy - Archive ouverte HAL
Article Dans Une Revue Cell Reports Methods Année : 2021

Super-resolved live-cell imaging using Random Illumination Microscopy

Thomas Mangeat
Simon Labouesse
  • Fonction : Auteur
Marc Allain
Awoke Negash
Claire Estibal
  • Fonction : Auteur
Sylvain Cantaloube
  • Fonction : Auteur
Christian Rouvière
  • Fonction : Auteur
Anne Sentenac
  • Fonction : Auteur
  • PersonId : 836364

Résumé

Current super-resolution microscopy (SRM) methods suffer from an intrinsic complexity that might curtail their routine use in cell biology. We describe here random illumination microscopy (RIM) for live-cell imaging at super-resolutions matching that of 3D structured illumination microscopy, in a robust fashion. Based on speckled illumination and statistical image reconstruction, easy to implement and user-friendly, RIM is unaffected by optical aberrations on the excitation side, linear to brightness, and compatible with multicolor live-cell imaging over extended periods of time. We illustrate the potential of RIM on diverse biological applications, from the mobility of proliferating cell nuclear antigen (PCNA) in U2OS cells and kinetochore dynamics in mitotic cells to the 3D motion of myosin minifilaments deep inside tissues. RIM's inherent simplicity and extended biological applicability, particularly for imaging at increased depths, could help make SRM accessible to biology laboratories.
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Dates et versions

hal-02564126 , version 1 (06-11-2020)

Identifiants

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Thomas Mangeat, Simon Labouesse, Marc Allain, Awoke Negash, Emmanuel Martin, et al.. Super-resolved live-cell imaging using Random Illumination Microscopy. Cell Reports Methods, 2021, 1 (1), pp.100009. ⟨10.1016/j.crmeth.2021.100009⟩. ⟨hal-02564126⟩
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