Label-free volumetric quantitative imaging of the human somatic cell division by hyperspectral coherent anti-Stokes Raman scattering

Karuna, Arnica, Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Wiltshire, Marie, Errington, Rachel ORCID: https://orcid.org/0000-0002-8016-4376, Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 and Langbein, Wolfgang ORCID: https://orcid.org/0000-0001-9786-1023 2019. Label-free volumetric quantitative imaging of the human somatic cell division by hyperspectral coherent anti-Stokes Raman scattering. Analytical Chemistry 91 (4) , pp. 2813-2821. 10.1021/acs.analchem.8b04706

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Abstract

Quantifying the chemical composition of unstained intact tissue and cellular samples with high spatio-temporal resolution in three dimensions would provide a step change in cell and tissue analytics critical to progress the field of cell biology. Label-free optical microscopy offers the required resolution and non-invasiveness, yet quantitative imaging with chemical specificity is a challenging endeavor. In this work, we show that hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy can be used to provide quantitative volumetric imaging of human osteosarcoma cells at various stages through cell division, a fundamental component of the cell cycle progress resulting in the segregation of cellular content to produce two progeny. We have developed and applied a quantitative data analysis method to produce volumetric three-dimensional images of the chemical composition of the dividing cell in terms of water, proteins, DNAP (a mixture of proteins and DNA, similar to chromatin), and lipids. We then used these images to determine the dry masses of the corresponding organic components. The attribution of proteins and DNAP components was validated using specific well-characterised fluorescent probes, by comparison with correlative two-photon fluorescence microscopy of DNA and mitochondria. Furthermore, we map the same chemical components under perturbed conditions, employing a drug that interferes directly with cell division (Taxol), showing its influence on cell organization and the masses of proteins, DNAP, and lipids.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences Medicine Physics and Astronomy
Additional Information:	This ACS article is provided under the terms of this ACS AuthorChoice/Editors’ Choice via Creative Commons CC-BY agreement.
Publisher:	American Chemical Society
ISSN:	0003-2700
Date of First Compliant Deposit:	10 January 2019
Date of Acceptance:	9 January 2019
Last Modified:	22 Apr 2024 18:40
URI:	https://orca.cardiff.ac.uk/id/eprint/118270

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