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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.
Nature Communications
|August 30, 2020
Summary
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.
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.
