Fig. 1: Left-handed speed-optimized DNA-PAINT optimization.
From: Left-handed DNA for efficient highly multiplexed imaging at single-protein resolution
a Illustration of 12-plex multiplexing capability with six right-handed (R1, R2, R3, R4, R5, R6) and six left-handed (L1, L2, L3, L4, L5, L6) sequences. b 12-plex DNA origami demonstration. Left shows an exemplary field-of-view with respective left-handed and right-handed 15 nm DNA origami grids highlighted. Right: Exemplary 15 nm DNA origami grid for each sequence. This data belongs to one dataset. c Sub-5 nm localization precision imaging in 3D of the three super-resolution benchmarking targets, nuclear pores (Nup96 - green - L2), tubulin filaments (α-tubulin - cyan - L1), and mitochondria (Tom20 - red - L4). Left: Overview of a single cell and a zoom-in with all proteins. Middle top: individual nuclear pore with single Nup96 copies, spaced ~14.3 nm apart (± 2 nm), resolved and a side view of the nuclear pore with the two rings spaced ~ 50 nm apart. Middle bottom: Exemplary 3D colored section of tubulin filaments with a zoom in into a cross-section of an individual tubulin filament of ~ 30 nm diameter. Right: Mitochondria with a cross-sectional side view, z-colored. d Kinetics characterization on DNA origami. The top violin plot shows the bright time (s) evaluation, while the bottom shows the dark time evaluation of all 12 sequences. Values are normalized to 100 pM imager concentration (e) Kinetics characterization in nuclear pores. The top violin plot shows the bright time (s) evaluation, while the bottom shows the dark time evaluation of all 12 sequences. Values are normalized to 100 pM imager concentration. In d and e, black lines indicate the minimum and maximum of the underlying data, black boxes indicate the two central quartiles, and the white line shows the median of the distribution. For details see Supplementary Figs. 2 and 3.
