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Resonant out-of-phase fluorescence microscopy and remote imaging overcome spectral limitations

Author

Listed:
  • Jérôme Quérard

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Ruikang Zhang

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Zsolt Kelemen

    (Universite Paris-Saclay)

  • Marie-Aude Plamont

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Xiaojiang Xie

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR
    Southern University of Science and Technology)

  • Raja Chouket

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Insa Roemgens

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Yulia Korepina

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Samantha Albright

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Eliane Ipendey

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Michel Volovitch

    (PSL Research University, Collège de France
    PSL Research University)

  • Hanna L. Sladitschek

    (European Molecular Biology Laboratory)

  • Pierre Neveu

    (European Molecular Biology Laboratory)

  • Lionel Gissot

    (Universite Paris-Saclay)

  • Arnaud Gautier

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Jean-Denis Faure

    (Universite Paris-Saclay)

  • Vincent Croquette

    (Laboratoire de Physique Statistique, CNRS, ENS)

  • Thomas Saux

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

  • Ludovic Jullien

    (UPMC Univ Paris 06, CNRS, PSL Research University
    Sorbonne Universités, UPMC Univ Paris 06, École Normale Supérieure, CNRS, PASTEUR)

Abstract

We present speed out-of-phase imaging after optical modulation (OPIOM), which exploits reversible photoswitchable fluorophores as fluorescent labels and combines optimized periodic illumination with phase-sensitive detection to specifically retrieve the label signal. Speed OPIOM can extract the fluorescence emission from a targeted label in the presence of spectrally interfering fluorophores and autofluorescence. Up to four fluorescent proteins exhibiting a similar green fluorescence have been distinguished in cells either sequentially or in parallel. Speed OPIOM is compatible with imaging biological processes in real time in live cells. Finally speed OPIOM is not limited to microscopy but is relevant for remote imaging as well, in particular, under ambient light. Thus, speed OPIOM has proved to enable fast and quantitative live microscopic and remote-multiplexed fluorescence imaging of biological samples while filtering out noise, interfering fluorophores, as well as ambient light.

Suggested Citation

  • Jérôme Quérard & Ruikang Zhang & Zsolt Kelemen & Marie-Aude Plamont & Xiaojiang Xie & Raja Chouket & Insa Roemgens & Yulia Korepina & Samantha Albright & Eliane Ipendey & Michel Volovitch & Hanna L. S, 2017. "Resonant out-of-phase fluorescence microscopy and remote imaging overcome spectral limitations," Nature Communications, Nature, vol. 8(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-00847-3
    DOI: 10.1038/s41467-017-00847-3
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