Dye-pair distance-distribution functions for tethered fluorophores as a function of d in aqueous (blue) and glycerol/water (gray) MD simulations. Results of MC SA calculations are shown for comparison in green. Post hoc statistical analysis using the ANOVA/Tukey method yields p < 0.05 for pairwise comparisons of all of the distributions in A and B. Based on this analysis, we can conclude that the differences in SA, MD (aqueous), and MD (glycerol/water) distance distributions for tethered ATTO dyes where d ≤ 4 nm are statistically significant. Differences in the distributions in C, where d = 7.3 nm, are not statistically significant, p = 0.48). (One column, 3.25″W × 6.95″H.) To see this figure in color, go online.