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Profiling pluripotent stem cells and organelles using synchrotron radiation infrared microspectroscopy
Author(s) -
Sandt Christophe,
Frederick Joni,
Dumas Paul
Publication year - 2013
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201200139
Subject(s) - sox2 , reprogramming , induced pluripotent stem cell , microbiology and biotechnology , golgi apparatus , organelle , chemistry , cytoplasm , homeobox protein nanog , stem cell , biology , endoplasmic reticulum , nucleus , cell , embryonic stem cell , biochemistry , gene
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. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)