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TECHNOLOGY DEVELOPMENT |
Modifications as a Marker of the Putative Stem Cell Region of Human Intestinal Crypts
aBiomedical Sciences Unit, Department of Biological Sciences, Lancaster University, Bailrigg, Lancaster, United Kingdom;
bCentre for Diabetes and Metabolic Medicine, Queen Mary's School of Medicine and Dentistry, Institute of Cell and Molecular Science, London, United Kingdom;
cDepartment of Physics, Lancaster University, Bailrigg, Lancaster, United Kingdom;
dDepartment of Gastroenterology and Hepatology, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taiwan;
eSynchrotron Radiation Department, Daresbury Laboratories, Science and Technologies Facilities Council, Warrington, United Kingdom;
fEuropean Synchrotron Radiation Facility, Grenoble, France;
gSharoe Green Unit, Lancashire Teaching Hospitals NHS Trust, Preston, United Kingdom
Key Words. Crypts • Fourier transform infrared microspectroscopy • Intestinal • Principal component analysis • Symmetric phosphate Synchrotron
Correspondence: Francis L. Martin, Ph.D., Biomedical Sciences Unit, Department of Biological Sciences, Lancaster University, Bailrigg, Lancaster LA1 4YQ, U.K. Telephone: +44-1524-594505; Fax: +44-1524-593192; e-mail: f.martin{at}lancaster.ac.uk
Received March 20, 2007;
accepted for publication September 18, 2007.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLS EXPRESS September 27, 2007.
Complex biomolecules absorb in the mid-infrared (
= 2–20 µm), giving vibrational spectra associated with structure and function. We used Fourier transform infrared (FTIR) microspectroscopy to "fingerprint" locations along the length of human small and large intestinal crypts. Paraffin-embedded slices of normal human gut were sectioned (10 µm thick) and mounted to facilitate infrared (IR) spectral analyses. IR spectra were collected using globar (15 µm x 15 µm aperture) FTIR microspectroscopy in reflection mode, synchrotron (
10 µm x 10 µm aperture) FTIR microspectroscopy in transmission mode or near-field photothermal microspectroscopy. Dependent on the location of crypt interrogation, clear differences in spectral characteristics were noted. Epithelial-cell IR spectra were subjected to principal component analysis to determine whether wavenumber-absorbance relationships expressed as single points in "hyperspace" might on the basis of multivariate distance reveal biophysical differences along the length of gut crypts. Following spectroscopic analysis, plotted clusters and their loadings plots pointed toward symmetric (
s)PO (1,080 cm–1) vibrations as a discriminating factor for the putative stem cell region; this proved to be a more robust marker than other phenotypic markers, such as β-catenin or CD133. This pattern was subsequently confirmed by image mapping and points to a novel approach of nondestructively identifying a tissue's stem cell location. sPO, probably associated with DNA conformational alterations, might facilitate a means of identifying stem cells, which may have utility in other tissues where the location of stem cells is unclear.
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