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

Author:

Murphy Neville12,Tipping William J.3ORCID,Braddick Henry J.4,Wilson Liam T.4,Tomkinson Nicholas C. O.4,Faulds Karen3,Graham Duncan3ORCID,Farràs Pau12

Affiliation:

1. School of Biological and Chemical Sciences University of Galway Galway H91CF50 Ireland

2. CÚRAM, The SFI Research Centre for Medical Devices University of Galway Galway H91 W2TY Ireland

3. Centre for Molecular Nanometrology WestCHEM Department of Pure and Applied Chemistry, Technology and Innovation Centre University of Strathclyde Glasgow G1 1RD United Kingdom

4. Department of Pure and Applied Chemistry University of Strathclyde Glasgow G1 1XL United Kingdom

Abstract

AbstractMultiplex 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.

Funder

Science Foundation Ireland

Engineering and Physical Sciences Research Council

European Regional Development Fund

Publisher

Wiley

Subject

General Chemistry,Catalysis

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