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Tracking single particles for hours via continuous DNA-mediated fluorophore exchange
Florian Stehr1, Johannes Stein1, Julian Bauer1
1Max Planck Institute of Biochemistry, Martinsried, Germany.
Nature Communications
|July 22, 2021
Summary
Continuous fluorophore exchange using DNA-PAINT overcomes photobleaching limitations in single-particle tracking. This method enables uninterrupted observation of biomolecules for extended durations, advancing biological process studies.
Area of Science:
- Biophysics
- Molecular Biology
- Nanotechnology
Background:
- Single-particle tracking commonly uses fixed dyes or fusion proteins, but photobleaching limits observation to seconds.
- This limitation restricts detailed statistical analysis and the study of rare biological events.
Purpose of the Study:
- To overcome photobleaching limitations in single-particle tracking.
- To enable uninterrupted, long-duration monitoring of single molecules.
Main Methods:
- Utilized DNA-PAINT for continuous fluorophore exchange.
- Attached single-stranded DNA handles to target molecules for reversible binding of fluorescent oligonucleotides.
Main Results:
- Achieved uninterrupted single-molecule monitoring for extended durations (tens of minutes).
- Successfully observed DNA origami structures on membranes over prolonged periods.
Conclusions:
- DNA-PAINT with continuous fluorophore exchange significantly extends observation times in single-particle tracking.
- This technique offers new possibilities for studying cellular processes on physiologically relevant timescales.
