Resolution doubling in light-sheet microscopy via oblique plane structured illumination

Bingying Chen, Bo Jui Chang, Philippe Roudot, Felix Zhou, Etai Sapoznik, Madeleine Marlar-Pavey, James B. Hayes, Peter T. Brown, Chih Wei Zeng, Talley Lambert, Jonathan R. Friedman, Chun Li Zhang, Dylan T. Burnette, Douglas P. Shepherd, Kevin M. Dean, Reto P. Fiolka

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Structured illumination microscopy (SIM) doubles the spatial resolution of a fluorescence microscope without requiring high laser powers or specialized fluorophores. However, the excitation of out-of-focus fluorescence can accelerate photobleaching and phototoxicity. In contrast, light-sheet fluorescence microscopy (LSFM) largely avoids exciting out-of-focus fluorescence, thereby enabling volumetric imaging with low photobleaching and intrinsic optical sectioning. Combining SIM with LSFM would enable gentle three-dimensional (3D) imaging at doubled resolution. However, multiple orientations of the illumination pattern, which are needed for isotropic resolution doubling in SIM, are challenging to implement in a light-sheet format. Here we show that multidirectional structured illumination can be implemented in oblique plane microscopy, an LSFM technique that uses a single objective for excitation and detection, in a straightforward manner. We demonstrate isotropic lateral resolution below 150 nm, combined with lower phototoxicity compared to traditional SIM systems and volumetric acquisition speed exceeding 1 Hz.

Original languageEnglish (US)
Pages (from-to)1419-1426
Number of pages8
JournalNature Methods
Issue number11
StatePublished - Nov 2022

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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