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Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation
Published on: October 15, 2019
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Fluorescence microscopy with 6 nm resolution on DNA origami
Mario Raab1, Jürgen J Schmied, Ija Jusuk
1Institute for Physical and Theoretical Chemistry and Braunschweig Integrated Centre of Systems Biology (BRICS), Braunschweig University of Technology, Hans-Sommer Str. 10, 38106 Braunschweig (Germany).
Summary
This study resolves two closely spaced DNA origami nanorulers using DNA PAINT superresolution microscopy. It demonstrates the technique
Area of Science:
- Optics and Photonics
- Nanotechnology
- Biophysics
Background:
- Superresolution microscopy resolution is often defined by point-spread-function width or localization accuracy.
- A key aspect of resolution is the ability to distinguish between two separate objects.
- DNA origami nanorulers offer a promising scaffold for precise nanoscale measurements.
Purpose of the Study:
- To address the discrepancy between single-object localization and the resolution of two distinct objects.
- To utilize DNA origami nanorulers for high-resolution imaging in superresolution microscopy.
- To quantitatively assess the resolution limits of DNA PAINT for closely spaced structures.
Main Methods:
- Employing DNA origami nanorulers with precisely defined docking site separations (18, 12, and 6 nm).
- Utilizing DNA PAINT (point accumulation for imaging in nanoscale topography) superresolution microscopy.
- Performing Monte Carlo simulations to analyze localization noise effects.
Main Results:
- Successfully resolved two docking sites on DNA origami nanorulers at distances of 18, 12, and 6 nm.
- Demonstrated the capability of DNA PAINT to resolve structures at the nanoscale.
- Identified and quantified the impact of localization noise on resolving very small distances.
Conclusions:
- DNA origami nanorulers are effective tools for validating superresolution microscopy resolution.
- DNA PAINT can resolve nanoscale features, but performance is influenced by localization accuracy.
- Understanding localization noise is crucial for interpreting results at the resolution limit.
