Determination of collagen fiber orientation in histological slides using Mueller microscopy and validation by second harmonic generation imaging

Stéphane Bancelin, André Nazac, Bicher Haj Ibrahim, Petr Dokládal, Etienne Decencière, Benjamin Teig, Huda Haddad, Hervé Fernandez, Marie-Claire Schanne-Klein, Antonello De Martino
Opt. Express. 2014-09-10; 22(19): 22561
DOI: 10.1364/oe.22.022561

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We studied the azimuthal orientations of collagen fibers in histological slides of uterine cervical tissue by two different microscopy techniques, namely Mueller polarimetry (MP) and Second Harmonic Generation (SHG). SHG provides direct visualization of the fibers with high specificity, which orientations is then obtained by suitable image processing. MP provides images of retardation (among other polarimetric parameters) due to the optical anisotropy of the fibers, which is enhanced by Picrosirius Red staining. The fiber orientations are then assumed to be those of the retardation slow axes. The two methods, though fully different from each other, provide quite similar maps of average fiber orientations. Overall, our results confirm that MP microscopy provides reliable images of dominant fiber orientations at a much lower cost that SHG, which remains the “gold standard” for specific imaging of collagen fibers using optical microscopy.

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