Supplementary Figure 3: Illustration of translational and rotational offsets between imaging planes, and integration of measured intensities from a point source in n different imaging channels.

a,b. Depiction of positions of three particles, i = 1, 2, 3, imaged in the reference channel at positions x_i,ref, y_i,ref (left) and separately in a compared channel n at positions x_i,n, y_i,n (right). In each case, the three positions defined the imaging plane. The offsets between each compared channel n and the reference channel was then defined by the translational offsets Δx, Δy (a) and the rotational angles α (around z-axis; a) and γ (around x-axis; b). c. Illustration of the translational and rotational operations relating the corresponding images of individual particles among the four channels. Simulated images of point sources, as described in d, on the EMCCD camera’s four sections (or dubbed channels in the main text) designated for I₀, I₄₅, I₉₀ and I₁₃₅ (left panel); the corresponding images in the four sections, identified on the basis of the translational and rotational operations, are indicated by numbers (right panel). d. Illustration of intensity integration with a simulated point source (left panel). The center of the point source was located at the pixel with the maximum intensity, which corresponded to the actual center here (middle panel). In the right panel, the area, labeled ‘I’, was designated to the data-capturing zone that was set to harbor ~90% of the total recorded intensity signal. This data-capturing zone was separated from the background zone (Bk) by a buffer zone (Bf). The total background intensity was obtained by integrating the pixels within the entire Bk zone.