IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v13y2022i1d10.1038_s41467-022-35333-y.html
   My bibliography  Save this article

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
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-022-35333-y
    File Function: Abstract
    Download Restriction: no

    File URL: https://libkey.io/10.1038/s41467-022-35333-y?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    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.
    Full references (including those not matched with items on IDEAS)

    Citations

    Citations are extracted by the CitEc Project, subscribe to its RSS feed for this item.
    as


    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.

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. Yilun Li & Fang Huang, 2024. "A statistical resolution measure of fluorescence microscopy with finite photons," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    2. 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.
    3. 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.
    4. 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.
    5. Chupao Lin & Juan Santo Domingo Peñaranda & Jolien Dendooven & Christophe Detavernier & David Schaubroeck & Nico Boon & Roel Baets & Nicolas Le Thomas, 2022. "UV photonic integrated circuits for far-field structured illumination autofluorescence microscopy," Nature Communications, Nature, vol. 13(1), pages 1-9, December.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-35333-y. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.