High-throughput multispot single-molecule spectroscopy

Ryan A. Colyer; Giuseppe Scalia; Taiho Kim; Ivan Rech; Daniele Resnati; Stefano Marangoni; Massimo Ghioni; Sergio Cova; Shimon Weiss; Xavier Michalet

doi:10.1117/12.841398

25 February 2010 High-throughput multispot single-molecule spectroscopy

Ryan A. Colyer, Giuseppe Scalia, Taiho Kim, Ivan Rech, Daniele Resnati, Stefano Marangoni, Massimo Ghioni, Sergio Cova, Shimon Weiss, Xavier Michalet

Author Affiliations +

Proceedings Volume 7571, Single Molecule Spectroscopy and Imaging III; 75710G (2010) https://doi.org/10.1117/12.841398
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

Solution-based single-molecule spectroscopy and fluorescence correlation spectroscopy (FCS) are powerful techniques to access a variety of molecular properties such as size, brightness, conformation, and binding constants. However, this is limited to low concentrations, which results in long acquisition times in order to achieve good statistical accuracy. Data can be acquired more quickly by using parallelization. We present a new approach using a multispot excitation and detection geometry made possible by the combination of three powerful new technologies: (i) a liquid crystal spatial light modulator to produce multiple diffraction-limited excitation spots; (ii) a multipixel detector array matching the excitation pattern and (iii) a low-cost reconfigurable multichannel counting board. We demonstrate the capabilities of this technique by reporting FCS measurements of various calibrated samples as well as single-molecule burst measurements.

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Ryan A. Colyer, Giuseppe Scalia, Taiho Kim, Ivan Rech, Daniele Resnati, Stefano Marangoni, Massimo Ghioni, Sergio Cova, Shimon Weiss, and Xavier Michalet "High-throughput multispot single-molecule spectroscopy", Proc. SPIE 7571, Single Molecule Spectroscopy and Imaging III, 75710G (25 February 2010); https://doi.org/10.1117/12.841398

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