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      An Ectopic Imaging Window for Intravital Imaging of Engineered Bone Tissue

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          Tissue engineering is a promising branch of regenerative medicine, but its clinical application remains limited because thorough knowledge of the in vivo repair processes in these engineered implants is limited. Common techniques to study the different phases of bone repair in mice are destructive and thus not optimal to gain insight into the dynamics of this process. Instead, multiphoton‐intravital microscopy (MP‐IVM) allows visualization of (sub)cellular processes at high resolution and frequency over extended periods of time when combined with an imaging window that permits optical access to implants in vivo. In this study, we have developed and validated an ectopic imaging window that can be placed over a tissue‐engineered construct implanted in mice. This approach did not interfere with the biological processes of bone regeneration taking place in these implants, as evidenced by histological and micro–computed tomography (μCT)‐based comparison to control ectopic implants. The ectopic imaging window permitted tracking of individual cells over several days in vivo. Furthermore, the use of fluorescent reporters allowed visualization of the onset of angiogenesis and osteogenesis in these constructs. Taken together, this novel imaging window will facilitate further analysis of the spatiotemporal regulation of cellular processes in bone tissue–engineered implants and provides a powerful tool to enhance the therapeutic potential of bone tissue engineering. © 2017 The Authors JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

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          Most cited references 38

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          For the past 25 years NIH Image and ImageJ software have been pioneers as open tools for the analysis of scientific images. We discuss the origins, challenges and solutions of these two programs, and how their history can serve to advise and inform other software projects.
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            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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              Optical properties of biological tissues: a review.

              A review of reported tissue optical properties summarizes the wavelength-dependent behavior of scattering and absorption. Formulae are presented for generating the optical properties of a generic tissue with variable amounts of absorbing chromophores (blood, water, melanin, fat, yellow pigments) and a variable balance between small-scale scatterers and large-scale scatterers in the ultrastructures of cells and tissues.

                Author and article information

                [ 1 ] Laboratory of Clinical and Experimental Endocrinology Department of Chronic Diseases, Metabolism and Ageing KU Leuven Leuven Belgium
                [ 2 ] Prometheus Division of Skeletal Tissue Engineering KU Leuven Leuven Belgium
                Author notes
                [* ] Address correspondence to: Geert Carmeliet, MD, PhD, Laboratory of Clinical and Experimental Endocrinology, Herestraat 49 bus 902, 3000 Leuven, Belgium. E‐mail: geert.carmeliet@

                JBMR Plus
                JBMR Plus
                JBMR Plus
                John Wiley and Sons Inc. (Hoboken )
                31 January 2018
                March 2018
                : 2
                : 2 ( doiID: 10.1002/jbm4.v2.2 )
                : 92-102
                6124161 10.1002/jbm4.10028 JBM410028
                © 2017 The Authors JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

                This is an open access article under the terms of the License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                Figures: 5, Tables: 0, Pages: 11, Words: 6154
                Funded by: Scientific Research‐Flanders
                Award ID: G.0A7213 and G.096414
                Funded by: BOF‐KU Leuven GOA
                Award ID: 3M120209
                Funded by: Hercules Foundation
                Award ID: AKUL/11/033
                Original Article
                Original Articles
                Custom metadata
                March 2018
                Converter:WILEY_ML3GV2_TO_NLMPMC version:version= mode:remove_FC converted:05.09.2018


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