Figure 4.

Non-specific labeling in two-color dSTORM. (A) The bin size in radius (10, 20, and 30nm) is drawn at scale. Proteins (orange, <5 nm in diameter) can associate with a lipid (blue shading). Non-specific labeling (red X) occurs randomly and does not typically associate directly with the lipid or protein. When a radius of 5–10 nm is used, the random background is excluded from the measurement by virtue of proximity. The shorter the distance, the more background is excluded allowing pair correlation for low abundant proteins or those with a high background. (B) Example of precision measurements from a dSTORM super-resolution instrument (Vutara VXL). The cells were stained for GM1 and PIP₂ lipids. Radial precisions in 4 different cell types varied between 4 and 8 nm. Hence very little random overlap occurs due to precision above 10 nm. Taken from Yuan et al., 2022 with permission [7].