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Proceedings Paper

Single-molecule FRET experiments with a red-enhanced custom technology SPAD
Author(s): Francesco Panzeri; Antonino Ingargiola; Ron R. Lin; Niusha Sarkhosh; Angelo Gulinatti; Ivan Rech; Massimo Ghioni; Sergio Cova; Shimon Weiss; Xavier Michalet
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Paper Abstract

Single-molecule fluorescence spectroscopy of freely diffusing molecules in solution is a powerful tool used to investigate the properties of individual molecules. Single-Photon Avalanche Diodes (SPADs) are the detectors of choice for these applications. Recently a new type of SPAD detector was introduced, dubbed red-enhanced SPAD (RE-SPAD), with good sensitivity throughout the visible spectrum and with excellent timing performance. We report a characterization of this new detector for single-molecule fluorescence resonant energy transfer (smFRET) studies on freely diffusing molecules in a confocal geometry and alternating laser excitation (ALEX) scheme. We use a series of doubly-labeled DNA molecules with donor-to-acceptor distances covering the whole range of useful FRET values. Both intensity-based (μs-ALEX) and lifetime-based (ns-ALEX) measurements are presented and compared to identical measurements performed with standard thick SPADs. Our results demonstrate the great potential of this new detector for smFRET measurements and beyond.

Paper Details

Date Published: 22 February 2013
PDF: 11 pages
Proc. SPIE 8590, Single Molecule Spectroscopy and Superresolution Imaging VI, 85900D (22 February 2013); doi: 10.1117/12.2003187
Show Author Affiliations
Francesco Panzeri, Politecnico di Milano (Italy)
Antonino Ingargiola, Univ. of California, Los Angeles (United States)
Ron R. Lin, Univ. of California, Los Angeles (United States)
Niusha Sarkhosh, Univ. of California, Los Angeles (United States)
Angelo Gulinatti, Politecnico di Milano (Italy)
Ivan Rech, Politecnico di Milano (Italy)
Massimo Ghioni, Politecnico di Milano (Italy)
Sergio Cova, Politecnico di Milano (Italy)
Shimon Weiss, Univ. of California, Los Angeles (United States)
Xavier Michalet, Univ. of California, Los Angeles (United States)


Published in SPIE Proceedings Vol. 8590:
Single Molecule Spectroscopy and Superresolution Imaging VI
Jörg Enderlein; Ingo Gregor; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling, Editor(s)

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