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Structured illumination microscopy using a photonic chip

Abstract

Structured illumination microscopy (SIM) enables live-cell super-resolution imaging of subcellular structures at high speeds. At present, linear SIM uses free-space optics to illuminate the sample with the desired light patterns; however, such arrangements are prone to misalignment and add cost and complexity to the microscope. Here, we present an alternative photonic chip-based two-dimensional SIM approach (cSIM) in which the conventional glass sample slide in a microscope is replaced by a planar photonic chip that importantly both holds and illuminates the specimen. The photonic chip reduces the footprint of the light illumination path of SIM to around 4 × 4 cm2. An array of optical waveguides on the chip creates standing wave interference patterns at different angles, which illuminate the sample via evanescent fields. High-refractive-index silicon nitride waveguides allow a 2.3 times enhancement in imaging spatial resolution, exceeding the usual 2 times limit of SIM. In summary, cSIM offers a simple, stable and affordable approach for performing two-dimensional super-resolution imaging over a large field of view.

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Fig. 1: The concept.
Fig. 2: Waveguide designs for cSIM.
Fig. 3: cSIM imaging.
Fig. 4: cSIM imaging of biological specimens.
Fig. 5: cSIM with increased resolution.

Data availability

The data associated with the figures and Supplementary information can be obtained at https://github.com/UiTcSIM/cSIM.git.

Code availability

The SIM reconstructions in Figs. 3 and 4 and in Supplementary Fig. 11 were performed using FAIR SIM, an open source code available at https://github.com/fairSIM. The SIM reconstructions in Fig. 5 were performed using code provided by R. Heintzmann at the Friedrich-Schiller-University Jena, Germany.

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Acknowledgements

We thank R. Heintzmann and S. Das for help with the SIM reconstruction algorithms and design of the fibre-optical phase modulator, respectively. We also acknowledge C. I. Øie and D. Wolfson for help with biological specimens and T. Huser for fruitful conceptual discussions. This work was supported by the European Research Council (grant no. 336716 to B.S.A.).

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Authors

Contributions

B.S.A. supervised and conceived this project and provided funding for the project. Ø.I.H. built the set-up, performed the experiments and analysed the data. J.-C.T., F.T.D. and O.G.H. designed the waveguide chip and the mask for fabrication. All authors contributed to chip design. Ø.I.H., F.T.D. and J.-C.T. characterized the waveguides. F.T.D. and Ø.I.H. fabricated the on-chip thermo-optics for phase modulation. M.L. performed all the simulations of cSIM. Ø.I.H. and B.S.A. mainly wrote the manuscript and all authors commented on the manuscript.

Corresponding author

Correspondence to Balpreet Singh Ahluwalia.

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Competing interests

B.SA. and O.G.H. have applied for patent GB1705660.7 on SIM-on-chip. B.S.A. and Ø.I.H. are founders of Chip NanoImaging (https://www.chipnano.com/) that is commercializing SIM-on-chip technology. The other authors declare no competing interests.

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Supplementary Information

Supplementary Figs. 1–17 and Notes 1–6.

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Helle, Ø.I., Dullo, F.T., Lahrberg, M. et al. Structured illumination microscopy using a photonic chip. Nat. Photonics 14, 431–438 (2020). https://doi.org/10.1038/s41566-020-0620-2

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