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Related Experiment Video

Updated: Nov 26, 2025

Label-Free Imaging of Single Proteins Secreted from Living Cells via iSCAT Microscopy
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Label-free, mass-sensitive single-molecule imaging using interferometric scattering microscopy.

Nikolas Hundt1

  • 1Ludwig-Maximilians-Universität München, Department of Cellular Physiology, Biomedical Centre, Großhaderner Str. 9, 82152 Planegg/Martinsried, Germany.

Essays in Biochemistry
|December 9, 2020
PubMed
Summary

Interferometric scattering (iSCAT) microscopy enables label-free single-molecule imaging and mass photometry. This technique offers a powerful, simple method for quantitative analysis of biomolecular equilibria and cellular processes.

Keywords:
iSCATlight microscopylight scatteringmass photometrymolecular interactionsingle-molecule

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Area of Science:

  • Biophysics
  • Microscopy
  • Biochemistry

Background:

  • Single-molecule imaging traditionally relies on fluorescence labeling.
  • Fluorescence imaging faces challenges with background noise and requires labeling.

Purpose of the Study:

  • Introduce interferometric scattering (iSCAT) microscopy.
  • Highlight iSCAT's capabilities for label-free imaging and mass photometry.
  • Discuss iSCAT as a complementary technique in biochemistry.

Main Methods:

  • Utilizes light scattering for detection without fluorescent labels.
  • Leverages improved sensitivity and size-dependent scattering signals.
  • Applies mass photometry to determine molecular mass distributions.

Main Results:

  • Demonstrates label-free detection and imaging of single biomolecules.
  • Enables quantitative analysis of biomolecular equilibria with low sample consumption.
  • Facilitates quantitative measurements from single molecules to mesoscopic cellular dynamics.

Conclusions:

  • iSCAT microscopy is an emerging technology for label-free imaging.
  • Mass photometry via iSCAT provides quantitative insights into molecular mass.
  • Mass-sensitive iSCAT complements existing biochemical techniques effectively.