Analysis of fixed and live single cells using optical photothermal infrared with concomitant raman spectroscopy

Spadea, A; Denbigh, JL; Lawrence, MJ; Kansiz, M; Gardner, P

Abstract

This paper reports the first use of a novel completely optically based photothermal method (O-PTIR) for obtaining infrared spectra of both fixed and living cells using a quantum cascade laser (QCL) and optical parametric oscillator (OPO) laser as excitation sources, thus enabling all biologically relevant vibrations to be analyzed at submicron spatial resolution. In addition, infrared data acquisition is combined with concomitant Raman spectra from exactly the same excitation location, meaning the full vibrational profile of the cell can be obtained. The pancreatic cancer cell line MIA PaCa-2 and the breast cancer cell line MDA-MB-231 are used as model cells to demonstrate the capabilities of the new instrumentation. These combined modalities can be used to analyze subcellular structures in both fixed and, more importantly, live cells under aqueous conditions. We show that the protein secondary structure and lipid-rich bodies can be identified on the submicron scale.

Citation

Spadea, A., Denbigh, J., Lawrence, M., Kansiz, M., & Gardner, P. (2021). Analysis of fixed and live single cells using optical photothermal infrared with concomitant raman spectroscopy. Analytical Chemistry, 93(8), 3938-3950. https://doi.org/10.1021/acs.analchem.0c04846

• Journal Article Type: Article
• Acceptance Date: Jan 26, 2021
• Online Publication Date: Feb 17, 2021
• Publication Date: Mar 2, 2021
• Deposit Date: May 4, 2021
• Publicly Available Date: May 4, 2021
• Journal: Analytical Chemistry
• Print ISSN: 0003-2700
• Electronic ISSN: 1520-6882
• Publisher: American Chemical Society
• Volume: 93
• Issue: 8
• Pages: 3938-3950
• DOI: https://doi.org/10.1021/acs.analchem.0c04846
• Publisher URL: https://doi.org/10.1021/acs.analchem.0c04846
• Related Public URLs: http://pubs.acs.org/journal/iecac0
• Additional Information: Additional Information : ** Article version: VoR ** From Crossref journal articles via Jisc Publications Router ** Licence for VoR version of this article starting on 17-02-2021: https://creativecommons.org/licenses/by/4.0/ **Journal IDs: pissn 0003-2700; eissn 1520-6882 **History: published 02-03-2021; issued 17-02-2021; published_online 17-02-2021; Funders : AstraZeneca;University of Manchester;Division of Pharmacy and Optometry, University of Manchester; Projects : unspecified