Use of synchrotron-radiation-based FTIR imaging for characterizing changes in cell contents.
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 Si(3)N(4) 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 ν(C = C-H)/ν(as)(CH(3)) 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.
Université de Bordeaux, CNRS UMR 5248, Pessac, France.
Seydou
S
Yao
Michel
M
Moenner
Anders
A
Engdahl
Cyril
C
Petibois
D002460:Cell Line; D002470:Cell Survival; D042783:Endothelial Cells / Q000737:chemistry* / Q000166:cytology; D005227:Fatty Acids / Q000032:analysis; D006801:Humans; D055806:Microvessels / Q000166:cytology; D017433:Protein Structure, Secondary; D011506:Proteins / Q000032:analysis; D059629:Signal-To-Noise Ratio; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D017356:Synchrotrons
D005227:Fatty Acids; D011506:Proteins