Bessel-beam hyperspectral CARS microscopy with sparse sampling: enabling high-content high-throughput label-free quantitative chemical imaging

Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Pope, Iestyn ORCID: https://orcid.org/0000-0002-4104-0389, Watson, Peter ORCID: https://orcid.org/0000-0003-0250-7852, Langbein, Wolfgang ORCID: https://orcid.org/0000-0001-9786-1023 and Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 2018. Bessel-beam hyperspectral CARS microscopy with sparse sampling: enabling high-content high-throughput label-free quantitative chemical imaging. Analytical Chemistry 90 (6) , pp. 3775-3785. 10.1021/acs.analchem.7b04039

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Abstract

Microscopy-based high-content and high-throughput analysis of cellular systems plays a central role in drug discovery. However, for contrast and specificity, the majority of assays require a fluorescent readout which always comes with the risk of alteration of the true biological conditions. In this work, we demonstrate a label-free imaging platform which combines chemically specific hyperspectral coherent anti-Stokes Raman scattering microscopy with sparse sampling and Bessel beam illumination. This enabled us to screen multiwell plates at high speed, while retaining the high-content chemical analysis of hyperspectral imaging. To demonstrate the practical applicability of the method we addressed a critical side effect in drug screens, namely, drug-induced lipid storage within hepatic tissue. We screened 15 combinations of drugs and neutral lipids added to human HepG2 liver cells and developed a high-content quantitative data analysis pipeline which extracted the spectra and spatial distributions of lipid and protein components. We then used their combination to train a support vector machine discriminative algorithm. Classification of the drug responses in terms of phospholipidosis versus steatosis was achieved in a completely label-free assay.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Physics and Astronomy Biosciences
Publisher:	American Chemical Society
ISSN:	0003-2700
Funders:	Biotechnology and Biological Sciences Research Council
Date of First Compliant Deposit:	8 March 2018
Date of Acceptance:	5 March 2018
Last Modified:	15 Sep 2023 16:18
URI:	https://orca.cardiff.ac.uk/id/eprint/109746

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