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Identifying subpopulations in multicellular systems by quantitative chemical imaging using label-free hyperspectral CARS microscopy

Pope, Iestyn ORCID:, Masia, Francesco ORCID:, Ewan, Kenneth ORCID:, Jimenez-Pascual, Ana, Dale, Trevor C. ORCID:, Siebzehnrubl, Florian A. ORCID:, Borri, Paola ORCID: and Langbein, Wolfgang ORCID: 2021. Identifying subpopulations in multicellular systems by quantitative chemical imaging using label-free hyperspectral CARS microscopy. Analyst 146 (7) , pp. 2277-2291. 10.1039/D0AN02381G

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Quantitative hyperspectral coherent Raman scattering microscopy merges imaging with spectroscopy and utilises quantitative data analysis algorithms to extract physically meaningful chemical components, spectrally and spatially-resolved, with sub-cellular resolution. This label-free non-invasive method has the potential to significantly advance our understanding of the complexity of living multicellular systems. Here, we have applied an in-house developed hyperspectral coherent anti-Stokes Raman scattering (CARS) microscope, combined with a quantitative data analysis pipeline, to imaging living mouse liver organoids as well as fixed mouse brain tissue sections xenografted with glioblastoma cells. We show that the method is capable of discriminating different cellular sub-populations, on the basis of their chemical content which is obtained from an unsupervised analysis, i.e. without prior knowledge. Specifically, in the organoids, we identify sub-populations of cells at different phases in the cell cycle, while in the brain tissue, we distinguish normal tissue from cancer cells, and, notably, tumours derived from transplanted cancer stem cells versus non-stem glioblastoma cells. The ability of the method to identify different sub-populations was validated by correlative fluorescence microscopy using fluorescent protein markers. These examples expand the application portfolio of quantitative chemical imaging by hyperspectral CARS microscopy to multicellular systems of significant biomedical relevance, pointing the way to new opportunities in non-invasive disease diagnostics.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Physics and Astronomy
European Cancer Stem Cell Research Institute (ECSCRI)
Additional Information: This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.
Publisher: Royal Society of Chemistry
ISSN: 0003-2654
Funders: CRUK, MRC, Wellcome Trust
Date of First Compliant Deposit: 12 March 2021
Date of Acceptance: 10 February 2021
Last Modified: 11 Oct 2023 16:37

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