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Single-photon microscopy to study biomolecular condensates.

Eleonora Perego1, Sabrina Zappone1,2, Francesco Castagnetti3

  • 1Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia, Genoa, Italy.

Nature Communications
|December 12, 2023
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Summary
This summary is machine-generated.

Researchers developed a novel single-photon microscope for live-cell imaging. This advanced tool quantifies molecular dynamics within biomolecular condensates, offering new insights into cellular processes.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Molecular Imaging

Background:

  • Biomolecular condensates are essential membrane-less compartments regulating cellular processes.
  • Quantifying their molecular dynamics is challenging due to process diversity and condensate variability.
  • Conventional microscopy methods are insufficient for detailed dynamic analysis.

Purpose of the Study:

  • To present a novel single-photon microscope as a comprehensive framework for live-cell spectroscopy and imaging of biomolecular condensation.
  • To enhance quantitative confocal microscopy by accessing fluorescence signals at the single-photon level.

Main Methods:

  • Utilized a single-photon detector array for enhanced fluorescence signal detection.
  • Implemented photon spatiotemporal tagging for time-lapse super-resolved imaging.
  • Integrated fluorescence lifetime fluctuation spectroscopy for simultaneous monitoring of molecular mobility, interactions, and nano-environment properties.

Main Results:

  • Achieved time-lapse super-resolved imaging of molecular organization within sub-diffraction environments.
  • Simultaneously monitored molecular mobility, interactions, and nano-environment characteristics.
  • Revealed dynamics and interactions of RNA-binding proteins in stress granule formation across diverse scales.

Conclusions:

  • The developed single-photon microscope provides a versatile framework for investigating biomolecular condensation dynamics.
  • This platform enables correlative studies of molecular dynamics and nano-environment properties in live cells.
  • Opens new avenues for exploring diverse biomolecular processes beyond membrane-less organelles.