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      Noninvasive quantitative assessment of collagen degradation in parchments by polarization-resolved SHG microscopy

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          Abstract

          Conservation state of medieval parchments is noninvasively assessed by advanced quantitative nonlinear optical microscopy.

          Abstract

          Nondestructive and noninvasive investigation techniques are highly sought-after to establish the degradation state of historical parchments, which is up to now assessed by thermal techniques that are invasive and destructive. We show that advanced nonlinear optical (NLO) microscopy enables quantitative in situ mapping of parchment degradation at the micrometer scale. We introduce two parameters that are sensitive to different degradation stages: the ratio of two-photon excited fluorescence to second harmonic generation (SHG) signals probes severe degradation, while the anisotropy parameter extracted from polarization-resolved SHG measurements is sensitive to early degradation. This approach is first validated by comparing NLO quantitative parameters to thermal measurements on artificially altered contemporary parchments. We then analyze invaluable parchments from the Middle Ages and show that we can map their conservation state and assess the impact of a restoration process. NLO quantitative microscopy should therefore help to identify parchments most at risk and optimize restoration methods.

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          Most cited references39

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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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            Gelatin structure and composition linked to hard capsule dissolution: A review

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              Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms.

              Although the nonlinear optical effect known as second-harmonic generation (SHG) has been recognized since the earliest days of laser physics and was demonstrated through a microscope over 25 years ago, only in the past few years has it begun to emerge as a viable microscope imaging contrast mechanism for visualization of cell and tissue structure and function. Only small modifications are required to equip a standard laser-scanning two-photon microscope for second-harmonic imaging microscopy (SHIM). Recent studies of the three-dimensional in vivo structures of well-ordered protein assemblies, such as collagen, microtubules and muscle myosin, are beginning to establish SHIM as a nondestructive imaging modality that holds promise for both basic research and clinical pathology. Thus far the best signals have been obtained in a transmitted light geometry that precludes in vivo measurements on large living animals. This drawback may be addressed through improvements in the collection of SHG signals via an epi-illumination microscope configuration. In addition, SHG signals from certain membrane-bound dyes have been shown to be highly sensitive to membrane potential. Although this indicates that SHIM may become a valuable tool for probing cell physiology, the small signal size would limit the number of photons that could be collected during the course of a fast action potential. Better dyes and optimized microscope optics could ultimately lead to the imaging of neuronal electrical activity with SHIM.
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                Author and article information

                Journal
                Sci Adv
                Sci Adv
                SciAdv
                advances
                Science Advances
                American Association for the Advancement of Science
                2375-2548
                July 2021
                16 July 2021
                : 7
                : 29
                : eabg1090
                Affiliations
                [1 ]Laboratoire d’Optique et Biosciences, Ecole polytechnique, CNRS, INSERM, Institut Polytechnique de Paris, Palaiseau, France.
                [2 ]Centre de Recherche sur la Conservation (CRC), Muséum national d’Histoire naturelle, Ministère de la Culture, CNRS, Paris, France.
                [3 ]Université Paris-Saclay, Saint-Aubin, France.
                Author notes
                [* ]Corresponding author. Email: gael.latour@ 123456universite-paris-saclay.fr
                Author information
                http://orcid.org/0000-0002-0286-7850
                http://orcid.org/0000-0003-1326-2520
                http://orcid.org/0000-0002-6748-5378
                http://orcid.org/0000-0002-8337-3982
                http://orcid.org/0000-0002-8252-3707
                http://orcid.org/0000-0003-4452-1011
                http://orcid.org/0000-0003-3026-8932
                http://orcid.org/0000-0002-6244-6556
                Article
                abg1090
                10.1126/sciadv.abg1090
                8284887
                34272247
                97222cf4-cfdd-48de-9628-32e1b54228cb
                Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).

                This is an open-access article distributed under the terms of the Creative Commons Attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 16 December 2020
                : 02 June 2021
                Funding
                Funded by: doi http://dx.doi.org/10.13039/501100001665, Agence Nationale de la Recherche;
                Award ID: ANR-17-EURE-0021
                Funded by: doi http://dx.doi.org/10.13039/501100001665, Agence Nationale de la Recherche;
                Award ID: ANR-11-EQPX-0029
                Funded by: doi http://dx.doi.org/10.13039/501100001665, Agence Nationale de la Recherche;
                Award ID: ANR-10- INBS-04
                Categories
                Research Article
                Research Articles
                SciAdv r-articles
                Applied Physics
                Applied Sciences and Engineering
                Applied Physics
                Custom metadata
                Samantha Cecilio

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