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Use of synchrotron-radiation-based FTIR imaging for characterizing changes in cell contents

Yao, Seydou; Moenner, Michel; Engdahl, Anders LU and Petibois, Cyril (2012) In Analytical and Bioanalytical Chemistry 404(5). p.1311-1316
Abstract
FTIR imaging of individual cells is still limited by the low signal-to-noise ratio obtained from analysis of such weakly absorbing organic matter when using a Globar IR source. In this study, we used FTIR imaging with a synchrotron radiation source and a focal plane array detector to determine changes in the cellular contents of cryofixed cells after culture for 48 h on Si3N4 substrate. Several spectral differences were observed for cells deprived of glucose compared with control cells: a lower amide I-to-amide II ratio (P < 0.01); a different secondary structure profile of proteins (obtained from amide I spectral region curve fitting), with a significant increase in non-ordered structure components (P < 0.01); and a higher nu(C =... (More)
FTIR imaging of individual cells is still limited by the low signal-to-noise ratio obtained from analysis of such weakly absorbing organic matter when using a Globar IR source. In this study, we used FTIR imaging with a synchrotron radiation source and a focal plane array detector to determine changes in the cellular contents of cryofixed cells after culture for 48 h on Si3N4 substrate. Several spectral differences were observed for cells deprived of glucose compared with control cells: a lower amide I-to-amide II ratio (P < 0.01); a different secondary structure profile of proteins (obtained from amide I spectral region curve fitting), with a significant increase in non-ordered structure components (P < 0.01); and a higher nu(C = C-H)/nu (as)(CH3) absorption ratio (P < 0.01), suggesting increased unsaturation of fatty acyl chains. Therefore, our study has shown that FTIR imaging with a synchrotron radiation source enables determination of several spectral changes of individual cells between two experimental conditions, which thus opens the way to cell biology studies with this vibrational spectroscopy technique. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Bioanalytical methods, Spectroscopy/instrumentation, IR, spectroscopy/Raman spectroscopy, Imaging, Synchrotron radiation
in
Analytical and Bioanalytical Chemistry
volume
404
issue
5
pages
1311 - 1316
publisher
Springer
external identifiers
  • wos:000307957800006
  • scopus:84865474203
ISSN
1618-2642
DOI
10.1007/s00216-012-6223-0
language
English
LU publication?
yes
id
06afedc9-13f4-4215-b231-376b7f91661e (old id 3147600)
date added to LUP
2012-10-30 14:54:24
date last changed
2017-06-18 03:24:40
@article{06afedc9-13f4-4215-b231-376b7f91661e,
  abstract     = {FTIR imaging of individual cells is still limited by the low signal-to-noise ratio obtained from analysis of such weakly absorbing organic matter when using a Globar IR source. In this study, we used FTIR imaging with a synchrotron radiation source and a focal plane array detector to determine changes in the cellular contents of cryofixed cells after culture for 48 h on Si3N4 substrate. Several spectral differences were observed for cells deprived of glucose compared with control cells: a lower amide I-to-amide II ratio (P &lt; 0.01); a different secondary structure profile of proteins (obtained from amide I spectral region curve fitting), with a significant increase in non-ordered structure components (P &lt; 0.01); and a higher nu(C = C-H)/nu (as)(CH3) absorption ratio (P &lt; 0.01), suggesting increased unsaturation of fatty acyl chains. Therefore, our study has shown that FTIR imaging with a synchrotron radiation source enables determination of several spectral changes of individual cells between two experimental conditions, which thus opens the way to cell biology studies with this vibrational spectroscopy technique.},
  author       = {Yao, Seydou and Moenner, Michel and Engdahl, Anders and Petibois, Cyril},
  issn         = {1618-2642},
  keyword      = {Bioanalytical methods,Spectroscopy/instrumentation,IR,spectroscopy/Raman spectroscopy,Imaging,Synchrotron radiation},
  language     = {eng},
  number       = {5},
  pages        = {1311--1316},
  publisher    = {Springer},
  series       = {Analytical and Bioanalytical Chemistry},
  title        = {Use of synchrotron-radiation-based FTIR imaging for characterizing changes in cell contents},
  url          = {http://dx.doi.org/10.1007/s00216-012-6223-0},
  volume       = {404},
  year         = {2012},
}