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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10.1038/s41598-020-64891-8Licence: CC BY

Veettikazhy_2020_SR_Multi-photon_CC
Copyright © The Author(s) 2020. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
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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. (CoI)
1/08/17 → 31/07/22
Project: Standard
H2020 MSCA ITN - BE-OPTICAL: H2020 Marie Curie ITN 2015 BE-OPTICAL
Dholakia, K. (PI)
Joint Research Centre 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
Dataset

File

Wide-field multiphoton imaging through scattering media
Escobet Montalban, A. (Author), Dholakia, K. (Supervisor), 27 Jul 2020
Student thesis: Doctoral Thesis (PhD)

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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TY - JOUR

T1 - Multi-photon attenuation-compensated light-sheet fluorescence microscopy

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

ER -

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

Student theses

Wide-field multiphoton imaging through scattering media

Cite this