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Live single cell analysis using synchrotron FTIR microspectroscopy: development of a simple dynamic flow system for prolonged sample viability

Doherty, J; Raoof, A; Hussain, A; Wolna, M; Cinque, G; Brown, M; Gardner, P; Denbigh, JL

Live single cell analysis using synchrotron FTIR microspectroscopy: development of a simple dynamic flow system for prolonged sample viability Thumbnail


Authors

J Doherty

A Raoof

A Hussain

M Wolna

G Cinque

M Brown

P Gardner

JL Denbigh



Abstract

Synchrotron radiation Fourier transform infrared microspectroscopy (SR-microFTIR) of live biological cells has the potential to provide far greater biochemical and morphological detail than equivalent studies using dehydrated, chemically-fixed single cells. Attempts to measure live cells using microFTIR are complicated by the aqueous environment required and corresponding strong infrared absorbance by water. There is also the additional problem of the limited lifetime of the cells outside of their preferred culture environment. In this work, we outline simple, cost-effective modifications to a commercially available liquid sample holder to perform single live cell analysis under an IR microscope and demonstrate cell viability up to at least 24 hours. A study using this system in which live cells have been measured at increasing temperature has shown spectral changes in protein bands attributed to α-β transition, consistent with other published work, and proves the ability to simultaneously induce and measure biochemical changes. An additional study of deuterated palmitic acid (D31-PA) uptake at different timepoints has made use of over 200 individual IR spectra collected over ∼4 hours, taking advantage of the ability to maintain viable cell samples for longer periods of time in the measurement environment, and therefore acquire greatly increased numbers of spectra without compromising on spectral quality. Further developments of this system are planned to widen the range of possible experiments, and incorporate more complex studies, including drug-cell interaction.

Citation

Doherty, J., Raoof, A., Hussain, A., Wolna, M., Cinque, G., Brown, M., …Denbigh, J. (2019). Live single cell analysis using synchrotron FTIR microspectroscopy: development of a simple dynamic flow system for prolonged sample viability. Analyst, 144(3), 997-1007. https://doi.org/10.1039/C8AN01566J

Journal Article Type Article
Acceptance Date Oct 19, 2018
Online Publication Date Oct 31, 2018
Publication Date Feb 1, 2019
Deposit Date Jan 17, 2019
Publicly Available Date Oct 31, 2019
Journal Analyst
Print ISSN 0003-2654
Electronic ISSN 1364-5528
Publisher Royal Society of Chemistry
Volume 144
Issue 3
Pages 997-1007
DOI https://doi.org/10.1039/C8AN01566J
Publisher URL https://doi.org/10.1039/C8AN01566J
Related Public URLs https://pubs.rsc.org/en/journals/journalissues/an#!recentarticles

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