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Pulsed Interleaved MINFLUX

  • Luciano A. Masullo
    Luciano A. Masullo
    Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2390, C1425FQD, Ciudad Autónoma de Buenos Aires, Argentina
    Departamento de Física, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Güiraldes 2620, C1428EHA, Ciudad Autónoma de Buenos Aires, Argentina
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  • , 
  • Florian Steiner
    Florian Steiner
    Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13 Haus E, 81377 München, Germany
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  • , 
  • Jonas Zähringer
    Jonas Zähringer
    Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13 Haus E, 81377 München, Germany
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  • , 
  • Lucía F. Lopez
    Lucía F. Lopez
    Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2390, C1425FQD, Ciudad Autónoma de Buenos Aires, Argentina
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  • , 
  • Johann Bohlen
    Johann Bohlen
    Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13 Haus E, 81377 München, Germany
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  • , 
  • Lars Richter
    Lars Richter
    Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2390, C1425FQD, Ciudad Autónoma de Buenos Aires, Argentina
    Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13 Haus E, 81377 München, Germany
    More by Lars Richter
  • , 
  • Fiona Cole
    Fiona Cole
    Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13 Haus E, 81377 München, Germany
    More by Fiona Cole
  • , 
  • Philip Tinnefeld*
    Philip Tinnefeld
    Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13 Haus E, 81377 München, Germany
    *E-mail: [email protected]
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  • , and 
  • Fernando D. Stefani*
    Fernando D. Stefani
    Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2390, C1425FQD, Ciudad Autónoma de Buenos Aires, Argentina
    Departamento de Física, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Güiraldes 2620, C1428EHA, Ciudad Autónoma de Buenos Aires, Argentina
    *E-mail: [email protected]
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    Orcidhttp://orcid.org/0000-0002-3277-7215
Cite this: Nano Lett. 2021, 21, 1, 840–846
Publication Date (Web):December 18, 2020
https://doi.org/10.1021/acs.nanolett.0c04600
Copyright © 2020 American Chemical Society
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    Abstract

    Abstract Image

    We introduce p-MINFLUX, a new implementation of the highly photon-efficient single-molecule localization method with a simplified experimental setup and additional fluorescence lifetime information. In contrast to the original MINFLUX implementation, p-MINFLUX uses interleaved laser pulses to deliver the doughnut-shaped excitation foci at a maximum repetition rate. Using both static and dynamic DNA origami model systems, we demonstrate the performance of p-MINFLUX for single-molecule localization nanoscopy and tracking, respectively. p-MINFLUX delivers 1–2 nm localization precision with 2000–1000 photon counts. In addition, p-MINFLUX gives access to the fluorescence lifetime enabling multiplexing and super-resolved lifetime imaging. p-MINFLUX should help to unlock the full potential of innovative single-molecule localization schemes.

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.nanolett.0c04600.

    • Details about the optical setups, the system for drift correction, the preparation of DNA origami samples, measurement of the excitation beam pattern and the point-spread functions, p-MINFLUX measurement protocol, position estimator and Cramér-Rao bound, p-MINFLUX simulations, effect of misalignment of the excitation beam pattern, crosstalk between detection time windows, and simulations of time traces of the pointer origami. Also, it contains a list of oligonucleotides used for each DNA origami used (PDF)

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    Cited By

    This article is cited by 47 publications.

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