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• W2023285721 abstract "The quantification of collagen fibril size is a major issue for the investigation of pathological disorders associated with structural defects of the extracellular matrix. Second-harmonic generation microscopy is a powerful technique to characterize the macromolecular organization of collagen in unstained biological tissues. Nevertheless, due to the complex coherent building of this nonlinear optical signal, it has never been used to measure fibril diameter so far. Here we report absolute measurements of second-harmonic signals from isolated fibrils down to 30 nm diameter, via implementation of correlative second-harmonic-electron microscopy. Moreover, using analytical and numerical calculations, we demonstrate that the high sensitivity of this technique originates from the parallel alignment of collagen triple helices within fibrils and the subsequent constructive interferences of second-harmonic radiations. Finally, we use these absolute measurements as a calibration for ex vivo quantification of fibril diameter in the Descemet’s membrane of a diabetic rat cornea. Quantitative determination of collagen fibril size is important for investigating pathological disorders of the extracellular matrix. Here, the authors show that SHG microscopy is capable of measuring fibril diameter after calibration of SHG radiation using correlation to electron microscopy." @default.
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• W2023285721 date "2014-09-16" @default.
• W2023285721 modified "2023-09-30" @default.
• W2023285721 title "Determination of collagen fibril size via absolute measurements of second-harmonic generation signals" @default.
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• W2023285721 doi "https://doi.org/10.1038/ncomms5920" @default.
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• W2023285721 hasPublicationYear "2014" @default.
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