Increased optical pathlength through aqueous media for the infrared microanalysis of live cells

Doherty, J, Zhang, Z, Wehbe, K, Cinque, G, Gardner, P and Denbigh, JL ORCID: 2018, 'Increased optical pathlength through aqueous media for the infrared microanalysis of live cells' , Analytical and Bioanalytical Chemistry, 410 (23) , pp. 5779-5789.

PDF - Published Version
Available under License Creative Commons Attribution 4.0.

Download (2MB) | Preview


The study of live cells using Fourier transform infrared spectroscopy (FTIR) and FTIR microspectroscopy (FT-IRMS) intrinsically yields more information about cell metabolism than comparable experiments using dried or chemically fixed samples. There are, however, a number of barriers to obtaining high-quality vibrational spectra of live cells, including correction for the significant contributions of water bands to the spectra, and the physical stresses placed upon cells by compression in short pathlength sample holders. In this study, we present a water correction method that is able to result in good-quality cell spectra from water layers of 10 and 12 μm and demonstrate that sufficient biological detail is retained to separate spectra of live cells based upon their exposure to different novel anti-cancer agents. The IR brilliance of a synchrotron radiation (SR) source overcomes the problem of the strong water absorption and provides cell spectra with good signal-to-noise ratio for further analysis. Supervised multivariate analysis (MVA) and investigation of average spectra have shown significant separation between control cells and cells treated with the DNA cross-linker PL63 on the basis of phosphate and DNA-related signatures. Meanwhile, the same control cells can be significantly distinguished from cells treated with the protein kinase inhibitor YA1 based on changes in the amide II region. Each of these separations can be linked directly to the known biochemical mode of action of each agent.

Keywords: Synchrotron radiation (SR), Fourier transform infrared spectroscopy (FTIR), Infrared microspectroscopy (IRMS), Cancer, Single cell, Drug-cell interactions

Item Type: Article
Schools: Schools > School of Environment and Life Sciences > Biomedical Research Centre
Journal or Publication Title: Analytical and Bioanalytical Chemistry
Publisher: Springer
ISSN: 1618-2642
Related URLs:
Funders: Diamond Light Source
Depositing User: JL Denbigh
Date Deposited: 19 Sep 2018 13:41
Last Modified: 15 Feb 2022 23:51

Actions (login required)

Edit record (repository staff only) Edit record (repository staff only)


Downloads per month over past year