Expanding the range of bioorthogonal tags for multiplex stimulated Raman scattering microscopy

Neville Murphy; William J. Tipping; Henry J. Braddick; Liam T. Wilson; Nicholas C.O. Tomkinson; Karen Faulds; Duncan Graham; Pau Farràs

doi:10.1002/anie.202311530

Expanding the range of bioorthogonal tags for multiplex stimulated Raman scattering microscopy

Neville Murphy, William J. Tipping, Henry J. Braddick, Liam T. Wilson, Nicholas C.O. Tomkinson, Karen Faulds, Duncan Graham, Pau Farràs

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Abstract

Multiplex optical detection in live cells is challenging due to overlapping signals and poor signal-to-noise associated with some chemical reporters. To address this, the application of spectral phasor analysis to stimulated Raman scattering (SRS) microscopy for unmixing three bioorthogonal Raman probes within cells is reported. Triplex detection of a metallacarborane using the B−H stretch at 2480–2650 cm⁻¹, together with a bis-alkyne and deuterated fatty acid can be achieved within the cell-silent region of the Raman spectrum. When coupled to imaging in the high-wavenumber region of the cellular Raman spectrum, nine discrete regions of interest can be spectrally unmixed from the hyperspectral SRS dataset, demonstrating a new capability in the toolkit of multiplexed Raman imaging of live cells.

Original language	English
Article number	e202311530
Number of pages	8
Journal	Angewandte Chemie International Edition
Volume	62
Issue number	48
Early online date	27 Oct 2023
DOIs	https://doi.org/10.1002/anie.202311530
Publication status	Published - 27 Nov 2023

Keywords

stimulated Raman scattering
spectral phasor analysis
bioorthogonal labelling
imaging probes
Raman microscopy

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10.1002/anie.202311530Licence: CC BY 4.0

Murphy-etal-ACIE-2023-Expanding-the-range-of-bioorthogonal-tags-for-multiplex-stimulated-Raman-scattering-microscopyFinal published version, 4.08 MBLicence: CC BY 4.0

University Of Strathclyde - Equipment Account
Gachagan, A. (Principal Investigator), He, W. (Principal Investigator), Jaroszynski, D. (Principal Investigator), Martin, R. (Principal Investigator), McArthur, S. (Principal Investigator), McArthur, S. (Principal Investigator), Connolly, P. (Co-investigator), Edwards, P. (Co-investigator), Faulds, K. (Co-investigator), Florence, A. (Co-investigator), Graham, D. (Co-investigator), Leithead, B. (Co-investigator), Sefcik, J. (Co-investigator), Ter Horst, J. (Co-investigator), Trager-Cowan, C. (Co-investigator), Uttamchandani, D. (Co-investigator) & Wark, A. (Co-investigator)
EPSRC (Engineering and Physical Sciences Research Council)
1/08/13 → 28/02/23
Project: Research

Data for: "Expanding the Range of Bioorthogonal Tags for Multiplex Stimulated Raman Scattering Microscopy"
Tipping, W. (Creator), Murphy, N. (Contributor), Braddick, H. (Contributor), Wilson, L. T. (Contributor), Tomkinson, N. (Supervisor), Faulds, K. (Supervisor), Graham, D. (Supervisor) & Farras, P. (Supervisor), University of Strathclyde, 23 Oct 2023
DOI: 10.15129/0a0a96e1-6fd4-4e1e-856f-caa380bc1c14
Dataset

Cite this

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title = "Expanding the range of bioorthogonal tags for multiplex stimulated Raman scattering microscopy",

abstract = "Multiplex optical detection in live cells is challenging due to overlapping signals and poor signal-to-noise associated with some chemical reporters. To address this, the application of spectral phasor analysis to stimulated Raman scattering (SRS) microscopy for unmixing three bioorthogonal Raman probes within cells is reported. Triplex detection of a metallacarborane using the B−H stretch at 2480–2650 cm−1, together with a bis-alkyne and deuterated fatty acid can be achieved within the cell-silent region of the Raman spectrum. When coupled to imaging in the high-wavenumber region of the cellular Raman spectrum, nine discrete regions of interest can be spectrally unmixed from the hyperspectral SRS dataset, demonstrating a new capability in the toolkit of multiplexed Raman imaging of live cells.",

keywords = "stimulated Raman scattering, spectral phasor analysis, bioorthogonal labelling, imaging probes, Raman microscopy",

author = "Neville Murphy and Tipping, {William J.} and Braddick, {Henry J.} and Wilson, {Liam T.} and Tomkinson, {Nicholas C.O.} and Karen Faulds and Duncan Graham and Pau Farr{\`a}s",

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T1 - Expanding the range of bioorthogonal tags for multiplex stimulated Raman scattering microscopy

AU - Murphy, Neville

AU - Tipping, William J.

AU - Braddick, Henry J.

AU - Wilson, Liam T.

AU - Tomkinson, Nicholas C.O.

AU - Faulds, Karen

AU - Graham, Duncan

AU - Farràs, Pau

PY - 2023/11/27

Y1 - 2023/11/27

N2 - Multiplex optical detection in live cells is challenging due to overlapping signals and poor signal-to-noise associated with some chemical reporters. To address this, the application of spectral phasor analysis to stimulated Raman scattering (SRS) microscopy for unmixing three bioorthogonal Raman probes within cells is reported. Triplex detection of a metallacarborane using the B−H stretch at 2480–2650 cm−1, together with a bis-alkyne and deuterated fatty acid can be achieved within the cell-silent region of the Raman spectrum. When coupled to imaging in the high-wavenumber region of the cellular Raman spectrum, nine discrete regions of interest can be spectrally unmixed from the hyperspectral SRS dataset, demonstrating a new capability in the toolkit of multiplexed Raman imaging of live cells.

AB - Multiplex optical detection in live cells is challenging due to overlapping signals and poor signal-to-noise associated with some chemical reporters. To address this, the application of spectral phasor analysis to stimulated Raman scattering (SRS) microscopy for unmixing three bioorthogonal Raman probes within cells is reported. Triplex detection of a metallacarborane using the B−H stretch at 2480–2650 cm−1, together with a bis-alkyne and deuterated fatty acid can be achieved within the cell-silent region of the Raman spectrum. When coupled to imaging in the high-wavenumber region of the cellular Raman spectrum, nine discrete regions of interest can be spectrally unmixed from the hyperspectral SRS dataset, demonstrating a new capability in the toolkit of multiplexed Raman imaging of live cells.

KW - stimulated Raman scattering

KW - spectral phasor analysis

KW - bioorthogonal labelling

KW - imaging probes

KW - Raman microscopy

U2 - 10.1002/anie.202311530

DO - 10.1002/anie.202311530

M3 - Article

SN - 1433-7851

VL - 62

JO - Angewandte Chemie International Edition

JF - Angewandte Chemie International Edition

IS - 48

M1 - e202311530

ER -

Expanding the range of bioorthogonal tags for multiplex stimulated Raman scattering microscopy

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Data for: "Expanding the Range of Bioorthogonal Tags for Multiplex Stimulated Raman Scattering Microscopy"

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