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Chip-based wide field-of-view nanoscopy

Abstract

Present optical nanoscopy techniques use a complex microscope for imaging and a simple glass slide to hold the sample. Here, we demonstrate the inverse: the use of a complex, but mass-producible optical chip, which hosts the sample and provides a waveguide for the illumination source, and a standard low-cost microscope to acquire super-resolved images via two different approaches. Waveguides composed of a material with high refractive-index contrast provide a strong evanescent field that is used for single-molecule switching and fluorescence excitation, thus enabling chip-based single-molecule localization microscopy. Additionally, multimode interference patterns induce spatial fluorescence intensity variations that enable fluctuation-based super-resolution imaging. As chip-based nanoscopy separates the illumination and detection light paths, total-internal-reflection fluorescence excitation is possible over a large field of view, with up to 0.5 mm × 0.5 mm being demonstrated. Using multicolour chip-based nanoscopy, we visualize fenestrations in liver sinusoidal endothelial cells.

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Figure 1: Implementation of chip-based nanoscopy.
Figure 2: Demonstration of chip-based dSTORM.
Figure 3: Demonstration of chip-based ESI.
Figure 4: Imaging the same sample under varied acquisition conditions reveals the specific strengths of the different approaches.
Figure 5: Multi-colour chip-based dSTORM reveals the interplay between actin (magenta) and the membrane (green) in LSECs.

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Acknowledgements

The authors thank I. Yahiatène and M. Müller for help with the ESI reconstruction algorithm and V. Mönkemöller for help with sample preparation and membrane dye staining. This work was supported by the European Research Council (grant no. 336716 to B.S.A.), the Research Council of Norway (grant no. 244764/F11 to B.S.A.) and the German Academic Exchange Service (grant no. 57160327 to M.S.). R.D. acknowledges additional support from grant no. KF2140610NT4 of the German Federal Ministry for Economic Affairs and Energy.

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B.S.A. and M.S. conceived the project. All authors designed the research. C.I.Ø. isolated the cells and stained and prepared the biological samples. R.D. and Ø.I.H. built the set-up, prepared the non-biological samples, performed the experiments, performed the simulations, reconstructed the images, analysed the data and created the figures. R.D., Ø.I.H., M.S. and B.S.A. mainly wrote the paper. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Mark Schüttpelz or Balpreet S. Ahluwalia.

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Competing interests

M.S. and B.S.A. have applied for patent GB1606268.9 for chip-based optical nanoscopy. The other authors declare no competing financial interests.

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Diekmann, R., Helle, Ø., Øie, C. et al. Chip-based wide field-of-view nanoscopy. Nature Photon 11, 322–328 (2017). https://doi.org/10.1038/nphoton.2017.55

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