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Microscopy with self-reconstructing beams

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Abstract

Although self-reconstructing beams have been the focus of many scientific studies over the past decade, hardly anything is known about their propagation and self-healing behaviour in a three-dimensional, inhomogeneous medium. The controlled reduction of scattering and beam spreading would enable a new illumination concept for light microscopes, particularly for those designed to look deep into scattering tissue. By investigating three different classes of refractive index inhomogeneity, using two large glass spheres, a cluster of smaller spheres and a piece of human skin, respectively, we show that beam self-reconstruction is indeed possible. We demonstrate that a Bessel beam is unexpectedly robust against deflection at objects, and we define measures for self-reconstruction in this context. We present a prototype of a microscope with self-reconstructing beams (MISERB) and show that a holographically shaped, scanned Bessel beam not only reduces scattering artefacts, but also simultaneously increases image quality and penetration depth in dense media.

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Figure 1: Schematic of the set-up.
Figure 2: Simulation of intensity distributions of Gaussian and Bessel beams scattered by two spheres.
Figure 3: Measurement of beam propagation in a fluorescing gel with two large glass spheres arranged consecutively at various beam distances.
Figure 4: Images of a cluster of glass spheres.
Figure 5: Maximum-selection images of human skin.

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Acknowledgements

The authors thank Carl Zeiss MicroImaging GmbH (Jena, Germany) for support in instrument development. Further thanks go to Dr. M. Peschen and Dr. R. Pfister for providing fresh human skin as well as to Prof. Olaf Ronneberger, B. Bosworth, C. Gohn-Kreuz and P. v. Olshausen for helpful discussions. This study was supported by the Excellence Initiative of the German Federal and State Governments (EXC 294).

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F.F. designed the system, performed experiments and simulations, analysed the data and prepared all graphs. P.S. performed experiments on human skin. A.R. initiated and supervised the project, developed the theory and wrote the manuscript.

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Correspondence to Florian O. Fahrbach or Alexander Rohrbach.

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The authors declare no competing financial interests.

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Fahrbach, F., Simon, P. & Rohrbach, A. Microscopy with self-reconstructing beams. Nature Photon 4, 780–785 (2010). https://doi.org/10.1038/nphoton.2010.204

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