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Journal Article

Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT.

MPS-Authors
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Sahl,  S. J.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Hell,  S. W.       
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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2270432.pdf
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2270432_Suppl.pdf
(Supplementary material), 5MB

Citation

Hoyer, P., de Medeiros, G., Balazs, B., Norlin, N., Besir, C., Hanne, J., et al. (2016). Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT. Proceedings of the National Academy of Sciences of the United States of America, 113(13), 3442-3446. doi: 10.1073/pnas.1522292113.


Cite as: https://hdl.handle.net/11858/00-001M-0000-002A-35BB-1
Abstract
We present a plane-scanning RESOLFT [reversible saturable/switchable optical (fluorescence) transitions] light-sheet (LS) nanoscope, which fundamentally overcomes the diffraction barrier in the axial direction via confinement of the fluorescent molecular state to a sheet of subdiffraction thickness around the focal plane. To this end, reversibly switchable fluorophores located right above and below the focal plane are transferred to a nonfluorescent state at each scanning step. LS-RESOLFT nanoscopy offers wide-field 3D imaging of living biological specimens with low light dose and axial resolution far beyond the diffraction barrier. We demonstrate optical sections that are thinner by 5–12-fold compared with their conventional diffraction-limited LS analogs.