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Image Scanning Microscopy with Single-Photon Detector Array

Marco Castello, View ORCID ProfileGiorgio Tortarolo, View ORCID ProfileMauro Buttafava, View ORCID ProfileTakahiro Deguchi, View ORCID ProfileFederica Villa, View ORCID ProfileSami Koho, Paolo Bianchini, Colin J. R. Sheppard, View ORCID ProfileAlberto Diaspro, View ORCID ProfileAlberto Tosi, View ORCID ProfileGiuseppe Vicidomini
doi: https://doi.org/10.1101/335596
Marco Castello
1Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Giorgio Tortarolo
1Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
2Dipartimento di Informatiche, Bioingegneria, Robotica e Ingegneria dei Sistemi, University of Genoa, Via All’Opera Pia 13, 16145, Genoa, Italy
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Mauro Buttafava
3Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo Da Vinci 32, 20133, Milan, Italy
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Takahiro Deguchi
4Nanoscopy and NIC@IIT, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Federica Villa
3Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo Da Vinci 32, 20133, Milan, Italy
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Sami Koho
1Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Paolo Bianchini
4Nanoscopy and NIC@IIT, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Colin J. R. Sheppard
4Nanoscopy and NIC@IIT, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Alberto Diaspro
4Nanoscopy and NIC@IIT, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Alberto Tosi
3Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo Da Vinci 32, 20133, Milan, Italy
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Giuseppe Vicidomini
1Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy
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Abstract

Image scanning microscopy (ISM) improves the spatial resolution of conventional confocal laser-scanning microscopy (CLSM), but current implementations reduce versatility and restrict its combination with fluorescence spectroscopy techniques, such as fluorescence lifetime. Here, we describe a natural design of ISM based on a fast single-photon detector array, which allows straightforward upgrade of an existing confocal microscope, without compromising any of its functionalities. In contrast to all-optical ISM implementations, our approach provides access to the raw scanned images, opening the way to adaptive reconstruction methods, capable of considering different imaging conditions and distortions. We demonstrate its utility in the context of fluorescence lifetime, deep, multicolor and live-cell imaging. This implementation will pave the way for a transparent and massive transition from conventional CLSM to ISM.

confocal microscopy | time-resolved spectroscopy | image scanning microscopy | single-photon detector array

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Posted June 02, 2018.
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Image Scanning Microscopy with Single-Photon Detector Array
Marco Castello, Giorgio Tortarolo, Mauro Buttafava, Takahiro Deguchi, Federica Villa, Sami Koho, Paolo Bianchini, Colin J. R. Sheppard, Alberto Diaspro, Alberto Tosi, Giuseppe Vicidomini
bioRxiv 335596; doi: https://doi.org/10.1101/335596
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Image Scanning Microscopy with Single-Photon Detector Array
Marco Castello, Giorgio Tortarolo, Mauro Buttafava, Takahiro Deguchi, Federica Villa, Sami Koho, Paolo Bianchini, Colin J. R. Sheppard, Alberto Diaspro, Alberto Tosi, Giuseppe Vicidomini
bioRxiv 335596; doi: https://doi.org/10.1101/335596

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