Label-free and multimodal second harmonic generation light sheet microscopy
Label-free and multimodal second harmonic generation light sheet microscopy
Light sheet microscopy (LSM) has emerged as one of most profound three dimensional (3D) imaging tools in the life sciences over the last decade. However, LSM is currently performed with fluorescence detection on one- or multi-photon excitation. Label-free LSM imaging approaches have been rather limited. Second Harmonic Generation (SHG) imaging is a label-free technique that has enabled detailed investigation of collagenous structures, including its distribution and remodelling in cancers and respiratory tissue, and how these link to disease. SHG is generally regarded as having only forward- and back-scattering components, apparently precluding the orthogonal detection geometry used in Light Sheet Microscopy. In this work we demonstrate SHG imaging on a light sheet microscope (SHG-LSM) using a rotated Airy beam configuration that demonstrates a powerful new approach to direct, without any further processing or deconvolution, 3D imaging of harmonophores such as collagen in biological samples. We provide unambiguous identification of SHG signals on the LSM through its wavelength and polarisation sensitivity. In a multimodal LSM setup we demonstrate that SHG and two-photon signals can be acquired on multiple types of different biological samples. We further show that SHG-LSM is sensitive to changes in collagen synthesis within lung fibroblast 3D cell cultures. This work expands on the existing optical methods available for use with light sheet microscopy, adding a further label-free imaging technique which can be combined with other detection modalities to realise a powerful multi-modal microscope for 3D bioimaging.
Hanrahan, Niall
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Lane, Simon I. R.
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Johnson, Peter
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Bourdakos, Konstantinos
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Brereton, Christopher
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Ridley, Robert A.
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Davies, Elizabeth R.
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Hosny, Neveen A.
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Spickermann, Gunnar
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Forster, Robert
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Malcolm, Graeme
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Davies, Donna
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Jones, Mark G.
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Mahajan, Sumeet
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Hanrahan, Niall
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Lane, Simon I. R.
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Johnson, Peter
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Bourdakos, Konstantinos
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Brereton, Christopher
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Ridley, Robert A.
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Davies, Elizabeth R.
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Hosny, Neveen A.
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Spickermann, Gunnar
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Forster, Robert
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Malcolm, Graeme
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Davies, Donna
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Jones, Mark G.
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Mahajan, Sumeet
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Hanrahan, Niall, Lane, Simon I. R., Johnson, Peter, Bourdakos, Konstantinos, Brereton, Christopher, Ridley, Robert A., Davies, Elizabeth R., Hosny, Neveen A., Spickermann, Gunnar, Forster, Robert, Malcolm, Graeme, Davies, Donna, Jones, Mark G. and Mahajan, Sumeet
(2020)
Label-free and multimodal second harmonic generation light sheet microscopy.
bioRxiv.
(doi:10.1101/2020.09.07.284703).
Abstract
Light sheet microscopy (LSM) has emerged as one of most profound three dimensional (3D) imaging tools in the life sciences over the last decade. However, LSM is currently performed with fluorescence detection on one- or multi-photon excitation. Label-free LSM imaging approaches have been rather limited. Second Harmonic Generation (SHG) imaging is a label-free technique that has enabled detailed investigation of collagenous structures, including its distribution and remodelling in cancers and respiratory tissue, and how these link to disease. SHG is generally regarded as having only forward- and back-scattering components, apparently precluding the orthogonal detection geometry used in Light Sheet Microscopy. In this work we demonstrate SHG imaging on a light sheet microscope (SHG-LSM) using a rotated Airy beam configuration that demonstrates a powerful new approach to direct, without any further processing or deconvolution, 3D imaging of harmonophores such as collagen in biological samples. We provide unambiguous identification of SHG signals on the LSM through its wavelength and polarisation sensitivity. In a multimodal LSM setup we demonstrate that SHG and two-photon signals can be acquired on multiple types of different biological samples. We further show that SHG-LSM is sensitive to changes in collagen synthesis within lung fibroblast 3D cell cultures. This work expands on the existing optical methods available for use with light sheet microscopy, adding a further label-free imaging technique which can be combined with other detection modalities to realise a powerful multi-modal microscope for 3D bioimaging.
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2020.09.07.284703v1.full
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e-pub ahead of print date: 8 September 2020
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Local EPrints ID: 472937
URI: http://eprints.soton.ac.uk/id/eprint/472937
PURE UUID: 6eb8e057-9dd3-4ad0-a0d1-2b32baa7ff19
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Date deposited: 06 Jan 2023 12:49
Last modified: 30 Nov 2024 03:05
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Contributors
Author:
Niall Hanrahan
Author:
Simon I. R. Lane
Author:
Peter Johnson
Author:
Konstantinos Bourdakos
Author:
Christopher Brereton
Author:
Robert A. Ridley
Author:
Elizabeth R. Davies
Author:
Neveen A. Hosny
Author:
Gunnar Spickermann
Author:
Robert Forster
Author:
Graeme Malcolm
Author:
Mark G. Jones
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