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

doi:10.1038/s41598-020-64891-8

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

Research output: Contribution to journal › Article › peer-review

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, can 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.

Original language	English
Article number	8090
Number of pages	10
Journal	Scientific Reports
Volume	10
DOIs	https://doi.org/10.1038/s41598-020-64891-8
Publication status	Published - 15 May 2020

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Veettikazhy_2020_SR_Multi-photon_CC
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Final published version, 3.46 MBLicence: CC BY

M Squared - USTAN Biophotonics Nexus: M Sqaured - St Andrews Biophotonics Nexus
Dholakia, K. (PI)
EPSRC
1/11/17 → 31/10/22
Project: Standard
Resonant and shaped photonics for under: Resonant and shaped photonics for understanding the physical and biomedical world
Dholakia, K. (PI) & Gather, M. C. (CoI)
1/08/17 → 31/07/22
Project: Standard
H2020 MSCA ITN - BE-OPTICAL: H2020 Marie Curie ITN 2015 BE-OPTICAL
Dholakia, K. (PI)
European Commission
1/10/15 → 30/09/19
Project: Standard

Data underpinning: Multi-photon attenuation-compensated light-sheet fluorescence microscopy
Veettikazhy, M. (Creator), Nylk, J. (Creator), Gasparoli, F. M. (Creator), Escobet Montalban, A. (Creator), Hansen, A. K. (Creator), Marti, D. (Creator), Andersen, P. E. (Creator) & Dholakia, K. (Creator), University of St Andrews, Oct 2019
DOI: 10.17630/5d8d3500-cd31-4776-8c01-57ad3507d7ff
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Cite this

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title = "Multi-photon attenuation-compensated light-sheet fluorescence microscopy",

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, can 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.",

author = "Madhu Veettikazhy and Jonathan Nylk and Federico Gasparoli and Adri{\`a} Escobet-Montalb{\'a}n and Hansen, {Anders Kragh} and Dominik Marti and Andersen, {Peter Eskil} and Kishan Dholakia",

note = "We thank the UK Engineering and Physical Sciences Research Council for funding (grants EP/P030017/1 and EP/R004854/1), the European Union{\textquoteright}s Horizon 2020 Framework Programme (H2020) (675512, BE-OPTICAL), the Danish Council for Independent Research (DFF FTP grant 7017-00021), and the Otto M{\o}nsted Foundation (grant 19-70-0109).",

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doi = "10.1038/s41598-020-64891-8",

language = "English",

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AU - Veettikazhy, Madhu

AU - Nylk, Jonathan

AU - Gasparoli, Federico

AU - Escobet-Montalbán, Adrià

AU - Hansen, Anders Kragh

AU - Marti, Dominik

AU - Andersen, Peter Eskil

AU - Dholakia, Kishan

N1 - We thank the UK Engineering and Physical Sciences Research Council for funding (grants EP/P030017/1 and EP/R004854/1), the European Union’s Horizon 2020 Framework Programme (H2020) (675512, BE-OPTICAL), the Danish Council for Independent Research (DFF FTP grant 7017-00021), and the Otto Mønsted Foundation (grant 19-70-0109).

PY - 2020/5/15

Y1 - 2020/5/15

N2 - 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, can 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.

AB - 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, can 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.

U2 - 10.1038/s41598-020-64891-8

DO - 10.1038/s41598-020-64891-8

M3 - Article

SN - 2045-2322

VL - 10

JO - Scientific Reports

JF - Scientific Reports

M1 - 8090

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Multi-photon attenuation-compensated light-sheet fluorescence microscopy

Abstract

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Projects

M Squared - USTAN Biophotonics Nexus: M Sqaured - St Andrews Biophotonics Nexus

Resonant and shaped photonics for under: Resonant and shaped photonics for understanding the physical and biomedical world

H2020 MSCA ITN - BE-OPTICAL: H2020 Marie Curie ITN 2015 BE-OPTICAL

Datasets

Data underpinning: Multi-photon attenuation-compensated light-sheet fluorescence microscopy

Cite this