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Multi-photon attenuation-compensated light-sheet fluorescence microscopy

Madhu Veettikazhy, View ORCID ProfileJonathan Nylk, Federico Gasparoli, Adrià Escobet-Montalbán, Anders Kragh Hansen, Dominik Marti, Peter Eskil Andersen, Kishan Dholakia
doi: https://doi.org/10.1101/2019.12.12.873877
Madhu Veettikazhy
1DTU Health Tech, Technical University of Denmark, Frederiksborgvej 399, 4000 Roskilde, Denmark
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  • For correspondence: madve@dtu.dk
Jonathan Nylk
2SUPA, School of Physics and Astronomy, University of St Andrews, North Haugh, St Andrews, KY16 9SS, UK
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  • ORCID record for Jonathan Nylk
Federico Gasparoli
2SUPA, School of Physics and Astronomy, University of St Andrews, North Haugh, St Andrews, KY16 9SS, UK
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Adrià Escobet-Montalbán
2SUPA, School of Physics and Astronomy, University of St Andrews, North Haugh, St Andrews, KY16 9SS, UK
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Anders Kragh Hansen
3DTU Fotonik, Technical University of Denmark, Frederiksborgvej 399, 4000 Roskilde, Denmark
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Dominik Marti
1DTU Health Tech, Technical University of Denmark, Frederiksborgvej 399, 4000 Roskilde, Denmark
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Peter Eskil Andersen
1DTU Health Tech, Technical University of Denmark, Frederiksborgvej 399, 4000 Roskilde, Denmark
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Kishan Dholakia
2SUPA, School of Physics and Astronomy, University of St Andrews, North Haugh, St Andrews, KY16 9SS, UK
4Department of Physics, College of Science, Yonsei University, Seoul 03722, South Korea
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Abstract

Attenuation of optical fields owing to scattering and absorption limits the penetration depth for imaging. Whilst aberration correction may be used, this is difficult to implement over a large field-of-view in heterogeneous tissue. Attenuation-compensation allows tailoring of the maximum lobe of a propagation-invariant light field and promises an increase in depth penetration for imaging. Here we show this promising approach may be implemented in multi-photon (two-photon) light-sheet fluorescence microscopy and, furthermore, be achieved in a facile manner utilizing a graded neutral density filter, circumventing the need for complex beam shaping apparatus. A “gold standard” system utilizing a spatial light modulator for beam shaping is used to benchmark our implementation. The approach will open up enhanced depth penetration in light-sheet imaging to a wide range of end users.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 12, 2019.
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Multi-photon attenuation-compensated light-sheet fluorescence microscopy
Madhu Veettikazhy, Jonathan Nylk, Federico Gasparoli, Adrià Escobet-Montalbán, Anders Kragh Hansen, Dominik Marti, Peter Eskil Andersen, Kishan Dholakia
bioRxiv 2019.12.12.873877; doi: https://doi.org/10.1101/2019.12.12.873877
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Multi-photon attenuation-compensated light-sheet fluorescence microscopy
Madhu Veettikazhy, Jonathan Nylk, Federico Gasparoli, Adrià Escobet-Montalbán, Anders Kragh Hansen, Dominik Marti, Peter Eskil Andersen, Kishan Dholakia
bioRxiv 2019.12.12.873877; doi: https://doi.org/10.1101/2019.12.12.873877

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