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Diffraction-unlimited three-dimensional optical nanoscopy with opposing lenses

Nature Photonics volume 3, pages 381–387 (2009)Cite this article

Abstract

The resolution of far-field optical microscopy stagnated for a century, but a quest began in the 1990s leading to nanoscale imaging of transparent fluorescent objects in three dimensions. Important elements in this pursuit were the synthesis of the aperture of two opposing lenses and the modulation or switching of the fluorescence of adjacent markers. The first element provided nearly isotropic three-dimensional resolution by improving the axial resolution by three- to sevenfold, and the second enabled the diffraction barrier to be overcome. Here, we review recent progress in the synergistic combination of these two elements which non-invasively provide an isotropic diffraction-unlimited three-dimensional resolution in transparent objects.

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Figure 1: Maximizing 3D resolution in far-field fluorescence nanoscopy by expanding the illumination and/or the detection aperture of the microscope with opposing lenses.
Figure 2: PSF shaping in a targeted fluorophore switching modality.
Figure 3: STED microscopy provides in situ access to 3D morphological information of nanostructured block copolymers.
Figure 4: Noninvasive 3D far-field fluorescence nanoscopy of the interior of mitochondria within integral cells.
Figure 5: 3D fluorescence nanoscopy with interferometric PALM.
Figure 6: 3D far-field fluorescence nanoscopy by STORM within mammalian (BS-C-1) cells.

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Acknowledgements

Substantial contributions to the isoSTED imaging applications reviewed in this paper are from Chaitanya Ullal (block co-polymers), as well as Christian Wurm and Stefan Jakobs (mitochondria). We also thank Andreas Schönle, Claas v. Middendorf, and Jan Keller for helpful discussions and Jaydev Jethwa for critical reading. This work was supported by grants of the Deutsche Forschungsgemeinschaft to A.E. and S.W.H (SFB 755).

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  1. Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, 37070, Germany

    Stefan W. Hell, Roman Schmidt & Alexander Egner

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  1. Stefan W. Hell
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Correspondence to Stefan W. Hell.

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Hell, S., Schmidt, R. & Egner, A. Diffraction-unlimited three-dimensional optical nanoscopy with opposing lenses. Nature Photon 3, 381–387 (2009). https://doi.org/10.1038/nphoton.2009.112

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