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      Advanced microscopy analysis of the micro-nanoscale architecture of human menisci

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          Abstract

          The complex inhomogeneous architecture of the human meniscal tissue at the micro and nano scale in the absence of artefacts introduced by sample treatments has not yet been fully revealed. The knowledge of the internal structure organization is essential to understand the mechanical functionality of the meniscus and its relationship with the tissue’s complex structure. In this work, we investigated human meniscal tissue structure using up-to-date non-invasive imaging techniques, based on multiphoton fluorescence and quantitative second harmonic generation microscopy complemented with Environmental Scanning Electron Microscopy measurements. Observations on 50 meniscal samples extracted from 6 human menisci (3 lateral and 3 medial) revealed fundamental features of structural morphology and allowed us to quantitatively describe the 3D organisation of elastin and collagen fibres bundles. 3D regular waves of collagen bundles are arranged in “honeycomb-like” cells that are comprised of pores surrounded by the collagen and elastin network at the micro-scale. This type of arrangement propagates from macro to the nanoscale.

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          Most cited references32

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          Fluorescence lifetime measurements and biological imaging.

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            Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation.

            Multicolor nonlinear microscopy of living tissue using two- and three-photon-excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intrinsic indicators within tissue, such as nicotinamide adenine dinucleotide, retinol, indoleamines, and collagen provides crucial information for physiology and pathology. The efficient application of multiphoton microscopy to intrinsic imaging requires knowledge of the nonlinear optical properties of specific cell and tissue components. Here we compile and demonstrate applications involving a range of intrinsic molecules and molecular assemblies that enable direct visualization of tissue morphology, cell metabolism, and disease states such as Alzheimer's disease and cancer.
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              Nonlinear magic: multiphoton microscopy in the biosciences.

              Multiphoton microscopy (MPM) has found a niche in the world of biological imaging as the best noninvasive means of fluorescence microscopy in tissue explants and living animals. Coupled with transgenic mouse models of disease and 'smart' genetically encoded fluorescent indicators, its use is now increasing exponentially. Properly applied, it is capable of measuring calcium transients 500 microm deep in a mouse brain, or quantifying blood flow by imaging shadows of blood cells as they race through capillaries. With the multitude of possibilities afforded by variations of nonlinear optics and localized photochemistry, it is possible to image collagen fibrils directly within tissue through nonlinear scattering, or release caged compounds in sub-femtoliter volumes.
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                Author and article information

                Contributors
                olga.barrera@ndorms.ox.ac.uk
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                10 December 2019
                10 December 2019
                2019
                : 9
                : 18732
                Affiliations
                [1 ]ISNI 0000 0004 1762 5517, GRID grid.10776.37, Università degli Studi di Palermo, ; Palermo, Italy
                [2 ]ISNI 0000 0004 1936 8948, GRID grid.4991.5, University of Oxford, ; Oxford, UK
                [3 ]IRCCS Istituto Ortopedico Rizzoli, Laboratorio di Biomeccanica e Innovazione Tecnologica, Bologna, Italy
                [4 ]ISNI 0000000417571846, GRID grid.7637.5, Università degli Studi of Brescia, ; Brescia, Italy
                [5 ]ISNI 0000 0001 2295 9843, GRID grid.16008.3f, University of Luxembourg, ; Luxembourg, Luxembourg
                [6 ]ISNI 0000 0001 0726 8331, GRID grid.7628.b, Oxford Brookes University, ; Oxford, UK
                Article
                55243
                10.1038/s41598-019-55243-2
                6904744
                31822796
                74aaf5a7-5286-4438-bc06-16867253ae6f
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 24 June 2019
                : 13 November 2019
                Categories
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                © The Author(s) 2019

                Uncategorized
                biophysics,anatomy
                Uncategorized
                biophysics, anatomy

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