Traditional diffraction-limited optical mapping has a minimum resolution of ~800 bases. We demonstrated super-localization of a target single-stranded DNA (ssDNA) with a dye-labeled probe ssDNA that had complementary sequence with a small part of the target DNA. Super-resolution imaging was achieved using a single-molecule localization method, motion blur point accumulation for imaging in nanoscale topography (mbPAINT). In mbPAINT, target ssDNA molecules were immobilized on the substrate, and the probe ssDNA diffusing in the bulk solution stochastically bound/unbound to the target ssDNA, which enabled super-localization of single probe ssDNA molecules. The locations of the probe molecules indicated the locations of the complementary target ssDNA with sequence specificity, which was confirmed via the use of a control, non-complementary probe. Resolution of ~30 nm was achieved, corresponding to a resolvable ssDNA length of ~50 bases.

Super-resolution mbPAINT for optical localization of single-stranded DNA

248th National Meeting (2014)

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