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

DOI: 10.1007/s00216-018-1188-2 DOI Help

Authors: James Doherty (University of Manchester; Diamond Light Source) , Zhe Zhang (University of Manchester) , Katia Wehbe (Diamond Light Source) , Gianfelice Cinque (Diamond Light Source) , Peter Gardner (University of Manchester) , Joanna Denbigh (University of Salford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Analytical And Bioanalytical Chemistry , VOL 99

State: Published (Approved)
Published: July 2018
Diamond Proposal Number(s): 15375 , 17811

Open Access Open Access

Abstract: 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.

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

Subject Areas: Biology and Bio-materials, Medicine


Instruments: B22-Multimode InfraRed imaging And Microspectroscopy

Documents:
10.1007_s00216-018-1188-2.pdf