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

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

17 Scopus citations


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.

Original languageEnglish
Title of host publicationSingle Molecule Spectroscopy and Imaging III
StatePublished - 2010
Externally publishedYes
EventSingle Molecule Spectroscopy and Imaging III - San Francisco, CA, United States
Duration: 23 Jan 201025 Jan 2010

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ISSN (Print)1605-7422


ConferenceSingle Molecule Spectroscopy and Imaging III
Country/TerritoryUnited States
CitySan Francisco, CA


  • CMOS
  • FCS
  • Fluorescence
  • High-throughput
  • LCOS
  • Photon counting
  • SPAD array
  • Single-molecule


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