title: Profiling pluripotent stem cells and organelles using synchrotron radiation infrared microspectroscopy.
abstract: FTIR micro-spectroscopy is a sensitive, non-destructive and label-free method offering diffraction-limited resolution with high signal-to-noise ratios when combined with a synchrotron radiation source. The vibrational signature of individual cells was used to validate an alternative strategy for reprogramming induced pluripotent stem cells generated from amniocytes. The iPSC lines PB09 and PB10, were reprogrammed from the same amniocyte cell line using respectively the Oct54, Sox2, Lin28, and Nanog and the Oct4 and Sox2 transcription factor cocktail. We show that cells reprogrammed by the two different sets of transfection factors have similar spectral signatures after reprogramming, except for a small subpopulation of cells in one of the cell lines. Mapping HeLa cells at subcellular resolution, we show that the Golgi apparatus, the cytoplasm and the nucleus have a specific spectral signature. The CH(3):CH(2) ratio is the highest in the nucleus and the lowest in the Golgi apparatus/endoplasmic reticulum, in agreement with the membrane composition of these organelles. This is confirmed by specific staining of the organelles with fluorescent dyes. Subcellular differentiation of cell compartments is also demonstrated in living cells.
authors.affiliation: Synchrotron SOLEIL, L'Orme des Merisiers, Gif-sur-Yvette, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Sandt
authors.forename: Joni
authors.initials: J
authors.lastname: Frederick
authors.forename: Paul
authors.initials: P
authors.lastname: Dumas
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D002467:Cell Nucleus / Q000378:metabolism; D065150:Cellular Reprogramming; D003593:Cytoplasm / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D044085:Microfluidics / Q000379:methods*; D015233:Models, Statistical; D015999:Multivariate Analysis; D013053:Spectrophotometry / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D017356:Synchrotrons; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D014157:Transcription Factors