16-Ch time-resolved single-molecule spectroscopy using line excitation

Antonino Ingargiola; Pietro Peronio; Eitan Lerner; Angelo Gulinatti; Ivan Rech; Massimo Ghioni; Shimon Weiss; Xavier Michalet

doi:10.1117/12.2256367

2 March 2017 16-Ch time-resolved single-molecule spectroscopy using line excitation

Antonino Ingargiola, Pietro Peronio, Eitan Lerner, Angelo Gulinatti, Ivan Rech, Massimo Ghioni, Shimon Weiss, Xavier Michalet

Author Affiliations +

Proceedings Volume 10071, Single Molecule Spectroscopy and Superresolution Imaging X; 100710Q (2017) https://doi.org/10.1117/12.2256367
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

Single-molecule spectroscopy on freely-diffusing molecules allows detecting conformational changes of biomolecules without perturbation from surface immobilization. Resolving fluorescence lifetimes increases the sensitivity in detecting conformational changes and overcomes artifacts common in intensity-based measurements. Common to all freely-diffusing techniques, however, are the long acquisition times. We report a time-resolved multispot system employing a 16-channel SPAD array and TCSPC electronics, which overcomes the throughput issue. Excitation is obtained by shaping a 532 nm pulsed laser into a line, matching the linear SPAD array geometry. We show that the line-excitation is a robust and cost-effective approach to implement multispot systems based on linear detector arrays.

Citation Download Citation

Antonino Ingargiola, Pietro Peronio, Eitan Lerner, Angelo Gulinatti, Ivan Rech, Massimo Ghioni, Shimon Weiss, and Xavier Michalet "16-Ch time-resolved single-molecule spectroscopy using line excitation", Proc. SPIE 10071, Single Molecule Spectroscopy and Superresolution Imaging X, 100710Q (2 March 2017); https://doi.org/10.1117/12.2256367

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