Jump to main content
Jump to site search


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

Author affiliations

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.

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

Back to tab navigation

Supplementary files

Publication details

The article was received on 13 Aug 2018, accepted on 19 Oct 2018 and first published on 31 Oct 2018


Article type: Paper
DOI: 10.1039/C8AN01566J
Citation: Analyst, 2018, Advance Article
  •   Request permissions

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

    J. Doherty, A. Raoof, A. Hussain, M. Wolna, G. Cinque, M. Brown, P. Gardner and J. Denbigh, Analyst, 2018, Advance Article , DOI: 10.1039/C8AN01566J

Search articles by author

Spotlight

Advertisements