Figure 2
Validation of SMLM counting strategy using monomeric and dimeric model proteins.
SMLM images of CD86-mEos2 (a) and CTLA-4-mEos2 (b) at the membrane of Hela cells. (c,e) Magnifications of region I or II boxed in (a) or (b) respectively. The inset shows a magnification of the boxed region. Distributions of Nblinks are generated from SMLM data sets. (d) For CD86-mEos2, the distribution is well approximated by a geometric distribution and yields pmembrane = 0.28 ± 0.01 as blink parameter, which is used as a fixed value for all other membrane proteins (n = 11 cells, adjusted R2 = 0.986 for CD86-mEos2). (f) For CTLA-4-mEos2, the distribution of Nblinks fits to a negative binomial distribution with an average number of N = 1.93 ± 0.02 underlying fluorescent probes per complex (n = 11 cells, adjusted R2 = 0.989 for CTLA-4-mEos2). Scale bars: 2 μm; inset 100 nm.
