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      Molecular contrast on phase-contrast microscope

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          Abstract

          An optical microscope enables image-based findings and diagnosis on microscopic targets, which is indispensable in many scientific, industrial and medical settings. A standard benchtop microscope platform, equipped with e.g., bright-field and phase-contrast modes, is of importance and convenience for various users because the wide-field and label-free properties allow for morphological imaging without the need for specific sample preparation. However, these microscopes never have capability of acquiring molecular contrast in a label-free manner. Here, we develop a simple add-on optical unit, comprising of an amplitude-modulated mid-infrared semiconductor laser, that is attached to a standard microscope platform to deliver the additional molecular contrast of the specimen on top of its conventional microscopic image, based on the principle of photothermal effect. We attach this unit, termed molecular-contrast unit, to a standard phase-contrast microscope, and demonstrate high-speed label-free molecular-contrast phase-contrast imaging of silica-polystyrene microbeads mixture and molecular-vibrational spectroscopic imaging of HeLa cells. Our simple molecular-contrast unit can empower existing standard microscopes and deliver a convenient accessibility to the molecular world.

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          Most cited references 27

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          Two-photon laser scanning fluorescence microscopy

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            Far-field optical nanoscopy.

             Stefan Hell (2007)
            In 1873, Ernst Abbe discovered what was to become a well-known paradigm: the inability of a lens-based optical microscope to discern details that are closer together than half of the wavelength of light. However, for its most popular imaging mode, fluorescence microscopy, the diffraction barrier is crumbling. Here, I discuss the physical concepts that have pushed fluorescence microscopy to the nanoscale, once the prerogative of electron and scanning probe microscopes. Initial applications indicate that emergent far-field optical nanoscopy will have a strong impact in the life sciences and in other areas benefiting from nanoscale visualization.
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              Scanning Confocal Optical Microscopy and Magnetic Resonance on Single Defect Centers

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                Author and article information

                Contributors
                ideguchi@gono.phys.s.u-tokyo.ac.jp
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                18 July 2019
                18 July 2019
                2019
                : 9
                Affiliations
                [1 ]ISNI 0000 0001 2151 536X, GRID grid.26999.3d, Department of Physics, , The University of Tokyo, ; Tokyo, 113-0033 Japan
                [2 ]ISNI 0000 0001 2151 536X, GRID grid.26999.3d, Department of Neurology, , The University of Tokyo, ; Tokyo, 113-0033 Japan
                [3 ]ISNI 0000 0004 0373 3971, GRID grid.136593.b, Graduate School of Information Science and Technology, , Osaka University, ; Osaka, 565-0871 Japan
                [4 ]ISNI 0000 0004 1754 9200, GRID grid.419082.6, PRESTO, Japan Science and Technology Agency, ; Saitama, 332-0012 Japan
                [5 ]ISNI 0000 0001 2151 536X, GRID grid.26999.3d, Institute for Photon Science and Technology, , The University of Tokyo, ; Tokyo, 113-0033 Japan
                Article
                46383
                10.1038/s41598-019-46383-6
                6637114
                31316091
                © 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/.

                Funding
                Funded by: FundRef https://doi.org/10.13039/501100009023, MEXT | JST | Precursory Research for Embryonic Science and Technology (PRESTO);
                Award ID: JPMJPR17G2
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100001691, MEXT | Japan Society for the Promotion of Science (JSPS);
                Award ID: 17H04852
                Award ID: 17K19071
                Award Recipient :
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                © The Author(s) 2019

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                phase-contrast microscopy, infrared spectroscopy

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