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Three-dimensional total internal reflection fluorescence nanoscopy with sub-10 nm resolution
Alan M. Szalai,
David J Williamson,
Dylan M Owen,
Fernando D. Stefani
Posted 05 Jul 2019
bioRxiv DOI: 10.1101/693994
Posted 05 Jul 2019
Here, we present a single-molecule localization microscopy (SMLM) analysis method that delivers sub-10 nm z-resolution when combined with 2D total internal reflection (TIR) fluorescence imaging via DNA point accumulation for imaging nanoscale topography (DNA-PAINT). Axial resolution is obtained from a precise measurement of the emission intensity of single molecules under evanescent field excitation. This method can be implemented on any conventional TIR wide-field microscope without modifications. We validate this approach by resolving the periodicity of alpha-tubulin assembly in microtubules, demonstrating isotropic resolution below 8 nm.
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