Single-molecule FRET experiments with a red-enhanced custom technology SPAD

Francesco Panzeri; Antonino Ingargiola; Ron R. Lin; Niusha Sarkhosh; Angelo Gulinatti; Ivan Rech; Massimo Ghioni; Sergio Cova; Shimon Weiss; Xavier Michalet

doi:10.1117/12.2003187

22 February 2013 Single-molecule FRET experiments with a red-enhanced custom technology SPAD

Francesco Panzeri, Antonino Ingargiola, Ron R. Lin, Niusha Sarkhosh, Angelo Gulinatti, Ivan Rech, Massimo Ghioni, Sergio Cova, Shimon Weiss, Xavier Michalet

Author Affiliations +

Proceedings Volume 8590, Single Molecule Spectroscopy and Superresolution Imaging VI; 85900D (2013) https://doi.org/10.1117/12.2003187
Event: SPIE BiOS, 2013, San Francisco, California, United States

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.

Citation Download Citation

Francesco Panzeri, Antonino Ingargiola, Ron R. Lin, Niusha Sarkhosh, Angelo Gulinatti, Ivan Rech, Massimo Ghioni, Sergio Cova, Shimon Weiss, and Xavier Michalet "Single-molecule FRET experiments with a red-enhanced custom technology SPAD", Proc. SPIE 8590, Single Molecule Spectroscopy and Superresolution Imaging VI, 85900D (22 February 2013); https://doi.org/10.1117/12.2003187

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available