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

ACS Appl Mater Interfaces. 2013 Oct 9;5(19):9338-43. doi: 10.1021/am403984k. Epub 2013 Sep 27.

Abstract

We demonstrate the application of superlocalization microscopy to identify sequence-specific portions of single-stranded DNA (ssDNA) with sequence resolution of 50 nucleotides, corresponding to a spatial resolution of 30 nm. Super-resolution imaging was achieved using a variation of a single-molecule localization method, termed as "motion blur" point accumulation for imaging in nanoscale topography (mbPAINT). The target ssDNA molecules were immobilized on the substrate. Short, dye-labeled, and complementary ssDNA molecules stochastically bound to the target ssDNA, with repeated binding events allowing super-resolution. Sequence specificity was demonstrated via the use of a control, noncomplementary probe. The results support the possibility of employing relatively inexpensive short ssDNAs to identify gene sequence specificity with improved resolution in comparison to the existing methods.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • DNA / chemistry
  • DNA / genetics
  • DNA / isolation & purification*
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / isolation & purification*
  • Microscopy
  • Nanotechnology*

Substances

  • DNA, Single-Stranded
  • DNA