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This study introduces novel reference materials and calibration methods for optical microscopy, achieving atomic-scale localization accuracy. This breakthrough addresses overconfidence in measurements and enhances nanoscale precision for critical applications.

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Area of Science:

  • Optical Microscopy
  • Nanotechnology
  • Metrology

Background:

  • Overconfidence in localization microscopy measurements is common due to uncalibrated systems and the assumption that precision limits accuracy.
  • Microscale systematic errors can invalidate nanoscale statistical uncertainties in localization microscopy.
  • Existing calibration methods for optical microscopes are often insufficient for achieving high accuracy.

Purpose of the Study:

  • To develop a comprehensive solution for accurate localization microscopy.
  • To create the first reference materials enabling atomic-scale localization errors.
  • To establish subnanometer localization accuracy in widefield optical microscopy.

Main Methods:

  • Development of subresolution aperture arrays as reference materials.
  • Novel calibration methods for microscope systems using aperture arrays.
  • Aberration corrections to match the precision limit of reference materials.
  • Correction and registration of multi-color localization data.

Main Results:

  • Achieved localization errors approaching the atomic scale across a submillimeter field.
  • Demonstrated general applicability of reference materials and calibration methods across different light sources.
  • Introduced critical-dimension localization microscopy for nanofabrication testing and quality control.
  • Applied reference materials to investigate the stability of fluorescent nanoparticles.

Conclusions:

  • Established a foundation for subnanometer localization accuracy in widefield optical microscopy.
  • Provided a comprehensive solution to overconfidence in localization microscopy measurements.
  • Enabled precise testing of nanofabrication processes and quality control of nanostructures.