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

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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 languageEnglish
Article numbere202311530
Number of pages8
JournalAngewandte Chemie International Edition
Issue number48
Early online date27 Oct 2023
Publication statusPublished - 27 Nov 2023


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


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