Super-resolved live-cell imaging using random illumination microscopy

Cell Rep Methods. 2021 Apr 30;1(1):100009. doi: 10.1016/j.crmeth.2021.100009. eCollection 2021 May 24.


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 S. pombe cells to the 3D motion of myosin minifilaments deep inside Drosophila tissues. RIM's inherent simplicity and extended biological applicability, particularly for imaging at increased depths, could help make SRM accessible to biology laboratories.

Keywords: aberration; computational imaging; fluorescence; live imaging; microscopy; speckle; super-resolution; variance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drosophila
  • Image Processing, Computer-Assisted*
  • Lighting*
  • Microscopy, Fluorescence / methods