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

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.

SUBMITTER: Hoyer P

PROVIDER: S-EPMC4822606 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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

Hoyer Patrick P de Medeiros Gustavo G Balázs Bálint B Norlin Nils N Besir Christina C Hanne Janina J Kräusslich Hans-Georg HG Engelhardt Johann J Sahl Steffen J SJ Hell Stefan W SW Hufnagel Lars L

Proceedings of the National Academy of Sciences of the United States of America 20160316 13

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 nanosc ...[more]

PMID: 26984498

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