Journal
NATURE METHODS
Volume 19, Issue 11, Pages 1419-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41592-022-01635-8
Keywords
-
Categories
Funding
- National Institutes of Health [1R01DK127589, 1R21HD105189, 5P30CA142543, 1RM1GM145399, U54CA268072, MIRA R35GM125028, R35GM133522, R35GM137894, 5R01NS117065]
- American Heart Association Graduate Student Fellowship [AHA 836090]
- Investissements d'Avenir French Government program [ANR-16-CONV-0001]
- Excellence Initiative of Aix-Marseille University: A*MIDEX
- Research Corporation for Science Advancement
- Frederick Gardner Cottrell Foundation [28041]
- Chan Zuckerberg Initiative [2021-236170(5022)]
- Scialog
Ask authors/readers for more resources
Structured illumination microscopy (SIM) improves the spatial resolution of fluorescence microscopy, while light-sheet fluorescence microscopy (LSFM) avoids exciting out-of-focus fluorescence. By combining SIM with LSFM using oblique plane microscopy, high-resolution three-dimensional imaging with low phototoxicity can be achieved.
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. The longstanding goal of combining the optical sectioning of light-sheet illumination and the resolving power of multidirectional structured illumination microscopy is realized using an oblique plane microscope for improved fast 3D subcellular imaging.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available