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Focus image scanning microscopy for sharp and gentle super-resolved microscopy

Author

Listed:
  • Giorgio Tortarolo

    (Istituto Italiano di Tecnologia
    Laboratory of Experimental Biophysics, EPFL)

  • Alessandro Zunino

    (Istituto Italiano di Tecnologia)

  • Francesco Fersini

    (Istituto Italiano di Tecnologia
    DIBRIS, University of Genoa)

  • Marco Castello

    (Istituto Italiano di Tecnologia
    Nanoscopy & NIC@IIT, Istituto Italiano di Tecnologia)

  • Simonluca Piazza

    (Istituto Italiano di Tecnologia
    Nanoscopy & NIC@IIT, Istituto Italiano di Tecnologia)

  • Colin J. R. Sheppard

    (Nanoscopy & NIC@IIT, Istituto Italiano di Tecnologia)

  • Paolo Bianchini

    (Nanoscopy & NIC@IIT, Istituto Italiano di Tecnologia)

  • Alberto Diaspro

    (Nanoscopy & NIC@IIT, Istituto Italiano di Tecnologia
    DIFI, University of Genoa)

  • Sami Koho

    (Istituto Italiano di Tecnologia)

  • Giuseppe Vicidomini

    (Istituto Italiano di Tecnologia)

Abstract

To date, the feasibility of super-resolution microscopy for imaging live and thick samples is still limited. Stimulated emission depletion (STED) microscopy requires high-intensity illumination to achieve sub-diffraction resolution, potentially introducing photodamage to live specimens. Moreover, the out-of-focus background may degrade the signal stemming from the focal plane. Here, we propose a new method to mitigate these limitations without drawbacks. First, we enhance a STED microscope with a detector array, enabling image scanning microscopy (ISM). Therefore, we implement STED-ISM, a method that exploits the working principle of ISM to reduce the depletion intensity and achieve a target resolution. Later, we develop Focus-ISM, a strategy to improve the optical sectioning and remove the background of any ISM-based imaging technique, with or without a STED beam. The proposed approach requires minimal architectural changes to a conventional microscope but provides substantial advantages for live and thick sample imaging.

Suggested Citation

  • 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.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-35333-y
    DOI: 10.1038/s41467-022-35333-y
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    References listed on IDEAS

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    1. 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.
    2. 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. 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.

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