Fig. 2: Resolution test and comparison to simulation.

a Tracking of beads fixed in agarose, the piezo stage is programmed to trace circles with an x, y-radius of 1 μm and a z peak-to-peak oscillation of 0.5 μm. Upper left; Trajectory estimated x (red), y (green) and z (blue) coordinates with a temporal resolution of 0.84 ms. Upper right; distribution of SD over the trajectory between estimation and a moving average with a window size of 100 points. Bottom left; photon counts per localization which corresponds to 60 kHz photon count rate. To the right is the photon count distribution. b Similar to a but for a simulated trajectory with an x,y-radius of 0.75 μm and a z peak-to-peak oscillation of 0.5 μm, the error distribution (upper right) is here the distance to ground truth. c Resolution as a function of photon count rate for x (red), y (green), and z (blue), solid lines are for the bead data, and dashed lines correspond to the simulations. Each point is the SD of the distance histogram in a and b. These, are obtained by increasing the exponential moving average of the likelihood. d 3D point scatter plot of the estimated (left) and simulated (right) trajectory. Source data are provided as a Source Data file.