Extended Data Fig. 6: Background localizations are imager-sequence specific.
From: Live-cell super-resolved PAINT imaging of piconewton cellular traction forces
a, Accumulated-tPAINT or (b) sf-tPAINT surfaces were incubated with scrambled-imager and specific imager. c, Representative images compiled from 2000 frames of single molecule localization for accumulated-tPAINT surfaces and (d) sf-tPAINT probes incubated with 5 nM scrambled and specific imagers. The average localizations (µm−2s−1) were quantified at 2.5, 5, and 10 nM scrambled and specific imagers for (e) accumulated-tPAINT (n = 4 independent experiments) and (f) sf-tPAINT surfaces (n = 3 independent experiments). Localization density was computed for 5 regions per image to compute an average (black points). Mean ± standard deviation is noted above each category. Error bars are 95% CI. 2.5 nM (accumulated-tPAINT, Scrambled): Mean= 0.0040, 95% CI 0.0015–0.0066; 2.5 nM (accumulated-tPAINT, Specific): Mean= 0.045, 95% CI −0.0017–0.092; 5 nM (accumulated-tPAINT, Scrambled): Mean= 0.0077, 95% CI −0.0057–0.02101; 5 nM (accumulated-tPAINT, Specific): Mean= 0.086, 95% CI −0.0032–0.17; 10 nM (accumulated-tPAINT, Scrambled): Mean= 0.0061, 95% CI 0.0025–0.0097; 10 nM (accumulated-tPAINT, Specific): Mean= 0.17, 95% CI −0.020–0.36; 2.5 nM (sf-tPAINT, Scrambled): Mean= 0.0024, 95% CI 0.00057–0.0043; 2.5 nM (sf-tPAINT, Specific): Mean= 0.022, 95% CI 0.0094–0.034; 5 nM (sf-tPAINT, Scrambled): Mean= 0.0035, 95% CI 0.0011–0.0059; 5 nM (sf-tPAINT, Specific): Mean= 0.028, 95% CI 0.0041–0.051; 10 nM (sf-tPAINT, Scrambled): Mean= 0.0037, 95% CI 0.0024–0.0050; 10 nM (sf-tPAINT, Specific): Mean= 0.044, 95% confidence interval −0.0048–0.094. Data were compared via a 2-way ANOVA. For localization on accumulated-tPAINT probe surface: 2.5 nM scramble versus 2.5 specific (p = 0.9968); 5 nM scramble versus 5 nM specific (p = 0.7527); 10 nM scramble is statistically different than 10 nM specific (p = 0.0115). For localization on sf-tPAINT probe surface: 2.5 nM scramble versus 2.5 specific (p = 0.3153); 5 nM scramble versus 5 nM specific (p = 0.1051); 10 nM scramble is statistically different than 10 nM specific (p = 0.0023).
