Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT
Abstract
Light-sheet fluorescence microscopy (LSFM) is an imaging modality in which a sample is illuminated from the side by a beam engineered into a wide and relatively thin "sheet." This allows highly parallelized planewise scanning of volumes with inherent optical sectioning, offering a good balance between spatial and temporal resolution with reduced photostress. Unfortunately, the axial extent of the illuminated section is ultimately limited by diffraction. Here, we show that a RESOLFT [reversible saturable/switchable optical (fluorescence) transitions] strategy neutralizes the resolution-limiting role of diffraction in LSFM. While other LS strategies exist, which run into new hard axial limits of resolution, LS-RESOLFT is conceptually diffraction-unlimited and can be developed toward molecular-scale resolution.
- Publication:
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Proceedings of the National Academy of Science
- Pub Date:
- March 2016
- DOI:
- 10.1073/pnas.1522292113
- Bibcode:
- 2016PNAS..113.3442H
- Keywords:
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- Physical Sciences,Applied Physical Sciences