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Updated: Jul 29, 2025

Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Ångström-resolution fluorescence microscopy.

Susanne C M Reinhardt1,2, Luciano A Masullo1, Isabelle Baudrexel1,3

  • 1Max Planck Institute of Biochemistry, Planegg, Germany.

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|May 24, 2023
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Summary

A new DNA-barcoding method, resolution enhancement by sequential imaging (RESI), achieves Ångström resolution with standard fluorescence microscopes. This breakthrough enables single-protein imaging in cells and mapping molecular arrangements for immunotherapy research.

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Microscopy

Background:

  • Fluorescence microscopy is crucial for life sciences but lacks resolution for molecular interactions.
  • Current super-resolution microscopy achieves ~5-20 nm resolution, insufficient for intramolecular structures.
  • Ångström resolution is needed for detailed molecular analysis but has not been achieved in cells.

Purpose of the Study:

  • To develop a method for achieving Ångström resolution using fluorescence microscopy.
  • To enable imaging of intramolecular structures and molecular arrangements in intact cells.
  • To provide a tool for understanding molecular mechanisms in biological systems and immunotherapy.

Main Methods:

  • Introduced resolution enhancement by sequential imaging (RESI), a DNA-barcoding technique.
  • Utilized sequential imaging of sparse target subsets at >15 nm resolution.
  • Employed off-the-shelf fluorescence microscopy hardware and reagents.

Main Results:

  • Achieved Ångström-scale resolution with standard fluorescence microscopy.
  • Demonstrated single-protein resolution in whole intact cells.
  • Resolved DNA backbone distances of single bases in DNA origami.
  • Mapped in situ molecular arrangement of CD20 in immunotherapy-relevant cells.

Conclusions:

  • RESI bridges the gap between super-resolution microscopy and structural biology.
  • Enables intramolecular imaging under ambient conditions in whole cells.
  • Offers new possibilities for studying molecular mechanisms in targeted immunotherapy.