Figure 2.

Assessment of the accuracy of subunit counting under different simulated experimental conditions. Measurement of the error against the (a) density of particles, (b) signal-to-noise ratio (SNR), (c) maximum number of subunits per complex, (d) number of monomeric particles selected for calibration, (e) intercomplex variation in photon count, (f) kernel probability distribution function (pdf) bin size, (g) ratio of the σ, or standard deviation, of the point spread function (PSF) of the underlying particles to ROI radius (SRR), and (h) pixel size. Base parameters used across all simulations (except for those varied): number of time frames, 500; number of movies for calibration and unknown species (each), at least 10; maximum photon count, 10; intercomplex variation in photon count, 0%; σ of the PSF of each complex, 130 nm; stoichiometry of the calibration species , 50% monomers, 25% dimers, and 25% trimers; stoichiometry of the unknown species, 50% monomers and 50% dimers. See the supplementary data for camera parameters.