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Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM
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Fast and multiplexed superresolution imaging with DNA-PAINT-ERS.

Fehmi Civitci1, Julia Shangguan2,3, Ting Zheng1

  • 1Knight Cancer Early Detection Advanced Research Center, Oregon Health and Science University, 2720 S. Moody Ave., Portland, OR, 97201, USA.

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We developed DNA-PAINT-ERS to speed up superresolution microscopy. This method enhances imaging speed and quality for DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) applications.

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Area of Science:

  • Biophysics
  • Microscopy
  • Molecular Biology

Background:

  • DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) enables multiplexed superresolution microscopy.
  • Current DNA-PAINT methods face limitations due to slow imaging speeds, hindering practical applications.

Purpose of the Study:

  • To enhance the imaging speed and quality of DNA-PAINT.
  • To introduce easily integrable modifications to existing DNA-PAINT workflows.

Main Methods:

  • Addition of ethylene carbonate (EC) to the imaging buffer.
  • Incorporation of repeating sequences into the docking strand.
  • Introduction of a spacer between the docking strand and affinity agent, collectively termed DNA-PAINT-ERS.

Main Results:

  • DNA-PAINT-ERS significantly accelerates imaging speed.
  • The modifications optimize DNA hybridization kinetics and efficiency, improving image quality.
  • Demonstrated general applicability across various DNA-PAINT setups with minor adjustments.

Conclusions:

  • DNA-PAINT-ERS offers a practical solution for faster, higher-quality superresolution imaging.
  • The strategy is broadly applicable and easily integrated into established DNA-PAINT protocols.
  • This advancement facilitates more efficient multiplexed superresolution microscopy.