Imaging experiments were performed using an inverted microscope (Ti Eclipse) with a 60×Plan-Apo (NA 1.40) objective lens (Nikon) and a spinning disk module (CSU-X1; Yokogawa). All images were acquired with Slidebook (3i – Intelligent Imaging Innovations) at room temperature. Images were captured with a charge-coupled device (CCD) camera (iXon X3; Andor Technology). Experiments were repeated 3 times and images were collected in three individual sets, as listed here for N = 1, 2, and 3.
Images are 3D stacks with only one channel, the resolution is 0.18 x 0.18 x 0.2 um^3, and stored in uint16 TIFF format.
Image processing was done on a desktop computer with a Core i9-10900K CPU @ 3.70GHz and 64 GB of RAM, running a Windows 11 operating system. The software was tested on MATLAB 2020b.