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.

The Characterisation of Pluripotent and Multipotent Stem Cells Using Fourier Transform Infrared Microspectroscopy

“…[38] compared the IR spectral profiles of two amniocyte derived IPSC lines that had been generated using different reprogramming. The iPSC line PB09 was transduced with four VSVG-pseudotyped lentiviruses carrying Oct4, Sox2, Lin28 and Nanog (OSLN) vectors.…”

Effects of Cannabis Use on the Protein and Lipid Profile of Olfactory Neuroepithelium Cells from Schizophrenia Patients Studied by Synchrotron-Based FTIR Spectroscopy

“…This non-destructive IR spectroscopy and imaging has a~1000-fold higher brightness than conventional global sources, thus, by improving the signal-to-noise ratio, it is very useful for the analysis of single cells [24]. Moreover, SR-FTIR is able to provide information regarding the quantity, composition, and distribution of chemical functional groups including nucleic acids, lipids and proteins in stain-free and label-free samples [25].…”

Re(I) carbonyl complexes: Multimodal platforms for inorganic chemical biology

“…In contrast, when vibrational levels are excited in the fundamental electronic state, the only dissipation is through the release of heat and thus no photobleaching occurs. In addition, IR microspectroscopy can be used to get chemical information about biological samples through signatures of lipids, esters, amides, or DNA [1][2][3]. The deeper penetration of IR radiation, in comparision with higher energy light, also enables tissues imaging.…”