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The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy

Author

Listed:
  • Alessandro Rossetta

    (Istituto Italiano di Tecnologia
    Istituto Italiano di Tecnologia
    University of Genoa)

  • Eli Slenders

    (Istituto Italiano di Tecnologia)

  • Mattia Donato

    (Istituto Italiano di Tecnologia)

  • Sabrina Zappone

    (Istituto Italiano di Tecnologia
    University of Genoa)

  • Francesco Fersini

    (Istituto Italiano di Tecnologia
    University of Genoa)

  • Martina Bruno

    (Istituto Italiano di Tecnologia
    University of Genoa)

  • Francesco Diotalevi

    (Istituto Italiano di Tecnologia)

  • Luca Lanzanò

    (Istituto Italiano di Tecnologia
    University of Catania)

  • Sami Koho

    (Istituto Italiano di Tecnologia)

  • Giorgio Tortarolo

    (Istituto Italiano di Tecnologia)

  • Andrea Barberis

    (Istituto Italiano di Tecnologia)

  • Marco Crepaldi

    (Istituto Italiano di Tecnologia)

  • Eleonora Perego

    (Istituto Italiano di Tecnologia)

  • Giuseppe Vicidomini

    (Istituto Italiano di Tecnologia)

Abstract

Fluorescence laser-scanning microscopy (LSM) is experiencing a revolution thanks to new single-photon (SP) array detectors, which give access to an entirely new set of single-photon information. Together with the blooming of new SP LSM techniques and the development of tailored SP array detectors, there is a growing need for (i) DAQ systems capable of handling the high-throughput and high-resolution photon information generated by these detectors, and (ii) incorporating these DAQ protocols in existing fluorescence LSMs. We developed an open-source, low-cost, multi-channel time-tagging module (TTM) based on a field-programmable gate array that can tag in parallel multiple single-photon events, with 30 ps precision, and multiple synchronisation events, with 4 ns precision. We use the TTM to demonstrate live-cell super-resolved fluorescence lifetime image scanning microscopy and fluorescence lifetime fluctuation spectroscopy. We expect that our BrightEyes-TTM will support the microscopy community in spreading SP-LSM in many life science laboratories.

Suggested Citation

  • Alessandro Rossetta & Eli Slenders & Mattia Donato & Sabrina Zappone & Francesco Fersini & Martina Bruno & Francesco Diotalevi & Luca Lanzanò & Sami Koho & Giorgio Tortarolo & Andrea Barberis & Marco , 2022. "The BrightEyes-TTM as an open-source time-tagging module for democratising single-photon microscopy," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-35064-0
    DOI: 10.1038/s41467-022-35064-0
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    References listed on IDEAS

    as
    1. Brian Owens, 2017. "The microscope makers," Nature, Nature, vol. 551(7682), pages 659-662, November.
    2. Enrica Maria Petrini & Tiziana Ravasenga & Torben J. Hausrat & Giuliano Iurilli & Umberto Olcese & Victor Racine & Jean-Baptiste Sibarita & Tija C. Jacob & Stephen J. Moss & Fabio Benfenati & Paolo Me, 2014. "Synaptic recruitment of gephyrin regulates surface GABAA receptor dynamics for the expression of inhibitory LTP," Nature Communications, Nature, vol. 5(1), pages 1-19, September.
    3. Sami Koho & Giorgio Tortarolo & Marco Castello & Takahiro Deguchi & Alberto Diaspro & Giuseppe Vicidomini, 2019. "Fourier ring correlation simplifies image restoration in fluorescence microscopy," Nature Communications, Nature, vol. 10(1), pages 1-9, December.
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    Cited by:

    1. Eleonora Perego & Sabrina Zappone & Francesco Castagnetti & Davide Mariani & Erika Vitiello & Jakob Rupert & Elsa Zacco & Gian Gaetano Tartaglia & Irene Bozzoni & Eli Slenders & Giuseppe Vicidomini, 2023. "Single-photon microscopy to study biomolecular condensates," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    2. Andrea Bucci & Giorgio Tortarolo & Marcus Oliver Held & Luca Bega & Eleonora Perego & Francesco Castagnetti & Irene Bozzoni & Eli Slenders & Giuseppe Vicidomini, 2024. "4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector," Nature Communications, Nature, vol. 15(1), pages 1-12, December.
    3. Giorgio Tortarolo & Alessandro Zunino & Francesco Fersini & Marco Castello & Simonluca Piazza & Colin J. R. Sheppard & Paolo Bianchini & Alberto Diaspro & Sami Koho & Giuseppe Vicidomini, 2022. "Focus image scanning microscopy for sharp and gentle super-resolved microscopy," Nature Communications, Nature, vol. 13(1), pages 1-14, December.

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