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Isolating stem cells in the inter-follicular epidermis employing synchrotron radiation-based Fourier-transform infrared microspectroscopy and focal plane array imaging

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Abstract

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.

“Delineating the putative stem cell lineage in interfollicular skin based on position-derived infrared spectral fingerprints”.

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Abbreviations

FPA:

Focal plane array

FTIR:

Fourier-transform infrared

H&E:

Haematoxylin and eosin

HCA:

Hierarchical cluster analysis

IR:

Infrared

LDA:

Linear discriminant analysis

NA:

Numerical aperture

PC:

Principal component

PCA:

Principal component analysis

SC:

stem cell

SNR:

Signal-to-noise ratio

TA:

Transit-amplifying

TD:

Terminally-differentiated

νasPO 2 :

Asymmetric phosphate stretching vibrations

νsPO 2 :

Symmetric phosphate stretching vibrations

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Acknowledgments

This work was funded by Rosemere Cancer Foundation and Unilever PLC. We also thanks the Sciences and Technologies Facilities Council for grant support to access the Diamond synchrotron facility.

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The authors indicate no potential conflicts of interest.

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Correspondence to Francis L. Martin.

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Patel, I.I., Harrison, W.J., Kerns, J.G. et al. Isolating stem cells in the inter-follicular epidermis employing synchrotron radiation-based Fourier-transform infrared microspectroscopy and focal plane array imaging. Anal Bioanal Chem 404, 1745–1758 (2012). https://doi.org/10.1007/s00216-012-6314-y

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  • DOI: https://doi.org/10.1007/s00216-012-6314-y

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