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Related Experiment Video

Updated: Mar 6, 2026

Label-Free Imaging of Single Proteins Secreted from Living Cells via iSCAT Microscopy
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Label-Free Single-Molecule Imaging with Numerical-Aperture-Shaped Interferometric Scattering Microscopy.

Daniel Cole1, Gavin Young1, Alexander Weigel1

  • 1Physical and Theoretical Chemistry Laboratory, University of Oxford , South Parks Road, OX1 3QZ Oxford, U.K.

ACS Photonics
|March 4, 2017
PubMed
Summary

A new optical microscopy technique uses a spatial mask to enhance extinction contrast, enabling label-free imaging of single molecules. This method improves the ability to optically interrogate nanoscopic objects.

Keywords:
biosensinginterferometric scatteringlabel-freesingle-molecule imaging

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Area of Science:

  • Optical microscopy
  • Nanotechnology
  • Biophysics

Background:

  • Interrogating nanoscopic objects optically is limited by extinction cross sections and diffraction.
  • Current methods struggle with low contrast for nanoscale imaging.

Purpose of the Study:

  • To enhance extinction contrast for improved optical interrogation of nanoscopic objects.
  • To enable routine label-free imaging at the single-molecule level.

Main Methods:

  • Utilizing a partially transmissive spatial mask near the back focal plane of a high numerical aperture microscope objective.
  • Employing numerical-aperture-based differentiation of background from scattered light.

Main Results:

  • Demonstrated an enhancement of extinction contrast by approximately T^-1/2, where T is the mask transmissivity.
  • Achieved routine label-free imaging down to the single-molecule level.

Conclusions:

  • The developed method provides a general approach to significantly increase extinction contrast.
  • This technique advances label-free nanoscale imaging capabilities.