File:TUD nanostructure.png

English: Single-molecule localization microscopy (SMLM) provides the ability to image well below the diffraction limit. The resolution in the final reconstructed image is limited by localization uncertainty and emitter density to about 20 nm. The fusion of multiple acquisitions into one hyper-resolved reconstruction can mitigate these limiting factors when many identical copies of the same structure (particle) can be imaged

We invented a template-free particle-fusion approach based on an all-to-all registration that provides robustness against individual misregistrations and underlabeling. We achieved 3.3-nm Fourier ring correlation (FRC) image resolution by fusing 383 DNA origami nanostructures with 80% labeling density.

The image shows the reconstruction from 383 individual super-resolution 2D PAINT images of DNA-origami structures of the TU Delft logo. We obtain wavelength/175 resolution. Each "blob" shows one binding site for fluorophores on the DNA-origmai structure. These sites are 5 nm apart (twice the width of DNA).

Image from our publication Heydarian et al. Nature Methods 2018:

https://doi.org/10.1038/s41592-018-0136-6