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Marker-free phase nanoscopy

Nature Photonics volume 7, pages 113–117 (2013)Cite this article

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A Corrigendum to this article was published on 29 April 2013

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Abstract

We introduce a microscopic method that determines quantitative optical properties beyond the optical diffraction limit and allows direct imaging of unstained living biological specimens. In established holographic microscopy, complex fields are measured using interferometric detection, allowing diffraction-limited phase measurements. Here, we show that non-invasive optical nanoscopy can achieve a lateral resolution of 90 nm by using a quasi-2π-holographic detection scheme and complex deconvolution. We record holograms from different illumination directions on the sample plane and observe subwavelength tomographic variations of the specimen. Nanoscale apertures serve to calibrate the tomographic reconstruction and to characterize the imaging system by means of the coherent transfer function. This gives rise to realistic inverse filtering and guarantees true complex field reconstruction. The observations are shown for nanoscopic porous cell frustule (diatoms), for the direct study of bacteria (Escherichia coli), and for a time-lapse approach to explore the dynamics of living dendritic spines (neurones).

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Figure 1: Marker-free nanoscopy by 2π-DHM in real and spatial frequency domains.
Figure 2: Side-by-side comparisons of resolution in traditional DHM and 2π-DHM.
Figure 3: Super-resolved phase images of nanoscopic porous cell frustule (diatoms).
Figure 4: Refraction tomogram of bacteria (E. coli).
Figure 5: Three-dimensional remodelling of a synaptic network from observations over a long time.

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  • 16 April 2013

    In the version of this Letter originally published online, no competing financial interests were declared. However, the authors wish to acknowledge a relevant patent. The competing financial interests statement has been modified in the HTML and PDF versions of the Letter.

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Acknowledgements

This work was funded by the Swiss National Science Foundation (SNSF, grant no. 205 320–130 543) and EPFL (innovation grant INNO 12-14). The authors acknowledge the Center of MicroNano-Technology (CMI) for cooperation regarding its research facilities. The authors give special thanks to M. Hildebrand for providing diatom samples, and to L. Pollaro for preparing the E. coli samples. The authors also thank P. Lara Rodrigo, C-M. Cotte and Cemico GmbH for assistance with graphics. Finally, the authors thank E. Cuche, CTO of Lyncée Tec, for helpful discussions and suggestions.

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Authors and Affiliations

  1. Advanced Photonics Laboratory, Microvision and Microdiagnostic Group (SCI STI CHD), Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, 1015, Switzerland

    Yann Cotte, Fatih Toy, Nicolas Pavillon, Daniel Boss & Christian Depeursinge

  2. Brain and Mind Institute, Laboratory of Neuroenergetics and Cellular Dynamics (LNDC), Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, 1015, Switzerland

    Pascal Jourdain, Daniel Boss, Pierre Magistretti & Pierre Marquet

  3. Department of Psychiatry-University Hospital, Centre de Neurosciences Psychiatriques, Prilly-Lausanne, 1008, Switzerland

    Pierre Magistretti & Pierre Marquet

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  1. Yann Cotte
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Contributions

Y.C., F.T., N.P. and C.D. designed the experiments. Y.C. and F.T. performed the experiments and carried out the main data analysis. P.J. and D.B. prepared the biological samples. C.D. and P.Mar. provided overall guidance to the project. All authors discussed the results and contributed to the manuscript.

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Correspondence to Yann Cotte.

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Competing interests

Yann Cotte, Nicolas Pavillon and Christian Depeursinge are named inventors on international patent WO/2011/121523 (publication date 06.10.2011, international filing date 28.03.2011), which is related to the techniques described in this Letter.

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Cotte, Y., Toy, F., Jourdain, P. et al. Marker-free phase nanoscopy. Nature Photon 7, 113–117 (2013). https://doi.org/10.1038/nphoton.2012.329

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