Super-resolution mbPAINT for optical localization of single-stranded DNA.
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.
Department of Chemistry and ‡Department of Electrical and Computer Engineering, Rice University , Houston, Texas 77251-1892, United States .
Jixin
J
Chen
Alberto
A
Bremauntz
Lydia
L
Kisley
Bo
B
Shuang
Christy F
CF
Landes
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000302:isolation & purification*; D008853:Microscopy; D036103:Nanotechnology
D004277:DNA, Single-Stranded; D004247:DNA