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Light-sheet microscopy using an Airy beam

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Abstract

Light-sheet microscopy facilitates rapid, high-contrast, volumetric imaging with minimal sample exposure. However, the rapid divergence of a traditional Gaussian light sheet restricts the field of view (FOV) that provides innate subcellular resolution. We show that the Airy beam innately yields high contrast and resolution up to a tenfold larger FOV. In contrast to the Bessel beam, which also provides an increased FOV, the Airy beam's characteristic asymmetric excitation pattern results in all fluorescence contributing positively to the contrast, enabling a step change for light-sheet microscopy.

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Figure 1: Airy beam light-sheet microscopy.
Figure 2: Experimental and simulated data for light-sheet illumination with various beam types.
Figure 3: Light-sheet microscopy scan of a juvenile amphioxus.

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  • 14 April 2014

    In the version of this article initially published online, reference 10 listed incorrect journal information: "Nat. Commun. 3, 632 (2012)" was erroneously listed as "Nat. Methods 9, 223 (2012)". The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

The project was supported by the UK Engineering and Physical Sciences Research Council, RS Macdonald Charitable Trust, Scottish Universities Life Sciences Alliance and St Andrews 600 anniversary BRAINS appeal. The PhD apprenticeship of C.C.L. was funded by the School of Biology. The authors thank A.M. Tilston-Lunel, M.L. Torres-Mapa, Z. Allen and A. Chatterton for their assistance with the cell culture; M. Ploschner for advice and proofreading of the manuscript; P.A. Reynolds for the fruitful discussions on the biological aspects of this study; and C. Penedo for his assistance with the photobleaching studies.

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Authors and Affiliations

Authors

Contributions

The Airy beam light sheet was developed by T.V. and K.D. T.V., H.I.C.D., T.Č. and J.N. conducted all the experiments. C.C.-L. and D.E.K.F. provided and prepared the amphioxus and zebrafish samples, and assisted with the imaging and interpretation. F.J.G.-M. assisted with all the biological experiments. J.N. and T.V. performed the simulations and data processing. T.V. and K.D. wrote the manuscript with contributions from all the authors.

Corresponding authors

Correspondence to Tom Vettenburg or Kishan Dholakia.

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Competing interests

T.V. and K.D. are authors on a patent application filed by the University Court of the University of St Andrews, UK, on this intellectual property right, which is currently progressing through the international patenting process. International Publication Number: WO 2013/150273 A1, published: 10 October 2013.

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Supplementary Figures 1–5 and Supplementary Notes 1–6 (PDF 5215 kb)

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Vettenburg, T., Dalgarno, H., Nylk, J. et al. Light-sheet microscopy using an Airy beam. Nat Methods 11, 541–544 (2014). https://doi.org/10.1038/nmeth.2922

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