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  A high-throughput image correlation method for rapid analysis of fluorophore photoblinking and photobleaching rates

Sehayek, S., Gidi, Y., Glembockyte, V., Brandão, H. B., François, P., Cosa, G., et al. (2019). A high-throughput image correlation method for rapid analysis of fluorophore photoblinking and photobleaching rates. ACS Nano, 13(10), 11955-11966. doi:10.1021/acsnano.9b06033.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000F-F7E8-5 Version Permalink: https://hdl.handle.net/21.11116/0000-000F-F7E9-4
Genre: Journal Article

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 Creators:
Sehayek, Simon, Author
Gidi, Yasser, Author
Glembockyte, Viktorija1, Author           
Brandão, Hugo B., Author
François, Paul, Author
Cosa, Gonzalo, Author
Wiseman, Paul W., Author
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1Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society, ou_2364732              

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 Abstract: Super-resolution fluorescence imaging based on localization microscopy requires tuning the photoblinking properties of fluorescent dyes employed. Missing is a rapid way to analyze the blinking rates of the fluorophore probes. Herein we present an ensemble autocorrelation technique for rapidly and simultaneously measuring photoblinking and bleaching rate constants from a microscopy image time series of fluorescent probes that is significantly faster than individual single-molecule trajectory analysis approaches. Our method is accurate for probe densities typically encountered in single-molecule studies as well as for higher density systems which cannot be analyzed by standard single-molecule techniques. We also show that we can resolve characteristic blinking times that are faster than camera detector exposure times, which cannot be accessed by threshold-based single-molecule approaches due to aliasing. We confirm this through computer simulation and single-molecule imaging data of DNA-Cy5 complexes. Finally, we demonstrate that with sufficient sampling our technique can accurately recover rates from stochastic optical reconstruction microscopy super-resolution data.

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Language(s): eng - English
 Dates: 2019-07-312019-09-122019-09-122019-10-22
 Publication Status: Issued
 Pages: 12
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1021/acsnano.9b06033
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Title: ACS Nano
  Abbreviation : ACS Nano
Source Genre: Journal
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Publ. Info: Washington, DC : American Chemical Society
Pages: - Volume / Issue: 13 (10) Sequence Number: - Start / End Page: 11955 - 11966 Identifier: ISSN: 1936-0851
CoNE: https://pure.mpg.de/cone/journals/resource/1936-0851