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

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

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.

Original languageEnglish
Title of host publicationSingle Molecule Spectroscopy and Superresolution Imaging X
EditorsZygmunt Karol Gryczynski, Jorg Enderlein, Ingo Gregor, Felix Koberling, Rainer Erdmann
PublisherSPIE
ISBN (Electronic)9781510605831
DOIs
StatePublished - 28 Jan 2017
Externally publishedYes
EventSingle Molecule Spectroscopy and Superresolution Imaging X - San Francisco, United States
Duration: 28 Jan 201729 Jan 2017

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume10071

Conference

ConferenceSingle Molecule Spectroscopy and Superresolution Imaging X
Country/TerritoryUnited States
CitySan Francisco
Period28/01/1729/01/17

Keywords

  • Fluorescence
  • High-throughput
  • SPAD array
  • Single-molecule
  • TCSPC

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Biomaterials

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