Although current implementations of super-resolution microscopy are technically approaching true molecular-scale resolution, this has not translated to imaging of biological specimens, because of the large size of conventional affinity reagents. Here we introduce slow off-rate modified aptamers (SOMAmers) as small and specific labeling reagents for use with DNA points accumulation in nanoscale topography (DNA-PAINT). To demonstrate the achievable resolution, specificity, and multiplexing capability of SOMAmers, we labeled and imaged both transmembrane and intracellular targets in fixed and live cells.
Optical
super-resolution techniques reach unprecedented spatial
resolution down to a few nanometers. However, efficient multiplexing
strategies for the simultaneous detection of hundreds of molecular
species are still elusive. Here, we introduce an entirely new approach
to multiplexed super-resolution microscopy by designing the blinking
behavior of targets with engineered binding frequency and duration
in DNA-PAINT. We assay this kinetic barcoding approach in
silico and in vitro using DNA origami structures,
show the applicability for multiplexed RNA and protein detection in
cells, and finally experimentally demonstrate 124-plex super-resolution
imaging within minutes.
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