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Precise Nanosizing with High Dynamic Range Holography.

Unai Ortiz-Orruño1, Ala Jo2,3, Hakho Lee2,3

  • 1ICFO, Institut de Ciencies Fotoniques, The Barcelona Institute of Science and Technology, Castelldefels 08860, Spain.

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Summary

This study introduces off-axis k-space holography, a novel optical sensing technique that overcomes dynamic range limitations for quantitative nanoparticle analysis. The method enables precise sizing of heterogeneous samples like extracellular vesicles.

Keywords:
Fourier imagingextracellular vesiclesholographyinterferometric microscopylabel-free

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Area of Science:

  • Biomedical Optics
  • Nanotechnology
  • Diagnostic Imaging

Background:

  • Label-free optical imaging is crucial for diagnostics, but challenges exist in quantifying heterogeneous samples like extracellular vesicles due to signal scaling issues.
  • Nanoparticle scattering signals vary with volume squared, exceeding dynamic range limits of current imaging cameras.
  • Quantitative characterization of intrinsically heterogeneous clinical samples remains a significant hurdle in optical sensing.

Purpose of the Study:

  • To introduce and validate a novel optical sensing technique, off-axis k-space holography, for overcoming dynamic range limitations.
  • To enable quantitative sizing and characterization of nanoparticles, including extracellular vesicles.
  • To demonstrate a platform for high-dynamic-range imaging of heterogeneous samples.

Main Methods:

  • Implementation of off-axis k-space holography by imaging the microscope's back-focal plane.
  • Projecting scattering signals of all particles onto all camera pixels to enhance dynamic range.
  • Utilizing independent signal calibrations for accurate sizing of particles from different materials.

Main Results:

  • Achieved a dramatically boosted dynamic range of up to 110 dB.
  • Successfully detected and quantitatively sized metallic and dielectric particles over a 200 × 200 μm field of view.
  • Presented quantitative size distributions for extracellular vesicle samples, demonstrating platform utility.

Conclusions:

  • Off-axis k-space holography effectively circumvents dynamic range limitations in optical sensing.
  • The validated platform enables accurate quantitative sizing of nanoparticles and heterogeneous clinical samples.
  • This technique holds promise for advancing diagnostics and nanoparticle characterization.