RT Journal Article
SR Electronic
T1 Image Scanning Microscopy with Single-Photon Detector Array
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 335596
DO 10.1101/335596
A1 Marco Castello
A1 Giorgio Tortarolo
A1 Mauro Buttafava
A1 Takahiro Deguchi
A1 Federica Villa
A1 Sami Koho
A1 Paolo Bianchini
A1 Colin J. R. Sheppard
A1 Alberto Diaspro
A1 Alberto Tosi
A1 Giuseppe Vicidomini
YR 2018
UL http://biorxiv.org/content/early/2018/06/02/335596.abstract
AB 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