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

Department of Physics

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

3 Citations (Scopus)

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
Journal	Scientific reports
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41598-020-64891-8
Publication status	Published - 2020 Dec 1

Bibliographical note

Funding Information:
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). We appreciate comments from G Spickermann (M Squared Lasers Ltd) on the manuscript. We thank Dr Wardiya Afshar Saber for providing the cell lines stably expressing GFP.

Publisher Copyright:
© 2020, The Author(s).

All Science Journal Classification (ASJC) codes

General

Access to Document

10.1038/s41598-020-64891-8

Cite this

@article{21b8323c7e9546edb4450079a042198f,

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 = "Funding Information: 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). We appreciate comments from G Spickermann (M Squared Lasers Ltd) on the manuscript. We thank Dr Wardiya Afshar Saber for providing the cell lines stably expressing GFP. Publisher Copyright: {\textcopyright} 2020, The Author(s).",

year = "2020",

month = dec,

day = "1",

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

language = "English",

volume = "10",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

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 - Funding Information: 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). We appreciate comments from G Spickermann (M Squared Lasers Ltd) on the manuscript. We thank Dr Wardiya Afshar Saber for providing the cell lines stably expressing GFP. Publisher Copyright: © 2020, The Author(s).

PY - 2020/12/1

Y1 - 2020/12/1

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.

UR - http://www.scopus.com/inward/record.url?scp=85084786005&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85084786005&partnerID=8YFLogxK

U2 - 10.1038/s41598-020-64891-8

DO - 10.1038/s41598-020-64891-8

M3 - Article

C2 - 32415135

AN - SCOPUS:85084786005

SN - 2045-2322

VL - 10

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 8090

ER -

Multi-photon attenuation-compensated light-sheet fluorescence microscopy

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this