Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - Isolating stem cells in the inter-follicular epidermis employing synchrotron radiation-based Fourier-transform infrared microspectroscopy and focal plane array imaging
AU - Patel, Imran I.
AU - Harrison, Wesley J.
AU - Kerns, Jemma G.
AU - Filik, Jacob
AU - Wehbe, Katia
AU - Carmichael, Paul L.
AU - Scott, Andrew D.
AU - Philpott, Mike P.
AU - Frogley, Mark D.
AU - Cinque, Gianfelice
AU - Martin, Francis L.
PY - 2012/10
Y1 - 2012/10
N2 - Normal function and physiology of the epidermis is maintained by the regenerative capacity of this tissue via adult stem cells (SCs). However, definitive identifying markers for SCs remain elusive. Infrared (IR) spectroscopy exploits the ability of cellular biomolecules to absorb in the mid-IR region (λ = 2.5-25 μm), detecting vibrational transitions of chemical bonds. In this study, we exploited the cell's inherent biochemical composition to discriminate SCs of the inter-follicular skin epidermis based on IR-derived markers. Paraffin-embedded samples of human scalp skin (n = 4) were obtained, and 10-μm thick sections were mounted for IR spectroscopy. Samples were interrogated in transmission mode using synchrotron radiation-based Fourier-transform IR (FTIR) microspectroscopy (15 × 15 μm) and also imaged employing globar-source FTIR focal plane array (FPA) imaging (5.4 × 5.4 μm). Dependent on the location of derived spectra, wavenumber-absorbance/intensity relationships were examined using unsupervised principal component analysis. This approach showed clear separation and spectral differences dependent on cell type. Spectral biomarkers concurrently associated with segregation of SCs, transit-amplifying cells and terminally-differentiated cells of epidermis were primarily PO(2)(-) vibrational modes (1,225 and 1,080 cm(-1)), related to DNA conformational alterations. FPA imaging coupled with hierarchical cluster analysis also indicated the presence of specific basal layer cells potentially originating from the follicular bulge, suggested by co-clustering of spectra. This study highlights PO (2) (-) vibrational modes as potential putative SC markers.
AB - Normal function and physiology of the epidermis is maintained by the regenerative capacity of this tissue via adult stem cells (SCs). However, definitive identifying markers for SCs remain elusive. Infrared (IR) spectroscopy exploits the ability of cellular biomolecules to absorb in the mid-IR region (λ = 2.5-25 μm), detecting vibrational transitions of chemical bonds. In this study, we exploited the cell's inherent biochemical composition to discriminate SCs of the inter-follicular skin epidermis based on IR-derived markers. Paraffin-embedded samples of human scalp skin (n = 4) were obtained, and 10-μm thick sections were mounted for IR spectroscopy. Samples were interrogated in transmission mode using synchrotron radiation-based Fourier-transform IR (FTIR) microspectroscopy (15 × 15 μm) and also imaged employing globar-source FTIR focal plane array (FPA) imaging (5.4 × 5.4 μm). Dependent on the location of derived spectra, wavenumber-absorbance/intensity relationships were examined using unsupervised principal component analysis. This approach showed clear separation and spectral differences dependent on cell type. Spectral biomarkers concurrently associated with segregation of SCs, transit-amplifying cells and terminally-differentiated cells of epidermis were primarily PO(2)(-) vibrational modes (1,225 and 1,080 cm(-1)), related to DNA conformational alterations. FPA imaging coupled with hierarchical cluster analysis also indicated the presence of specific basal layer cells potentially originating from the follicular bulge, suggested by co-clustering of spectra. This study highlights PO (2) (-) vibrational modes as potential putative SC markers.
KW - Biological Markers
KW - Cell Differentiation
KW - Epidermis
KW - Hair Follicle
KW - Humans
KW - Molecular Imaging
KW - Spectroscopy, Fourier Transform Infrared
KW - Stem Cells
U2 - 10.1007/s00216-012-6314-y
DO - 10.1007/s00216-012-6314-y
M3 - Journal article
C2 - 22945554
VL - 404
SP - 1745
EP - 1758
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
SN - 1618-2642
IS - 6-7
ER -