Automated Microscopy Imaging System for Cell Analysis

We have described TIS and the accompanying data analysis in previous publications^{29 (link),30} . The system used was the TIS basic 4 (pi4 Robotics GmbH, Berlin, Germany). It consists of a climate-controlled chamber containing: a Zeiss AxioImager microscope with a Colibri.2 lighting system and a Plan-Apochromat 63X/1.0 Ph3 M27 water immersion objective; an SC4022M digital imaging system (Finger Lakes Instrumentation, LLC, Lima, NY); and a robot-controlled motorized pipette. The TIS system included software programs (written by Reyk Hillert, Magdeburg, Germany) that were used to generate data for subsequent analysis. These included: Image Registrator v.1.1 (for image alignment and background subtraction); Binary Center v.1.0.2 (for binarization of images); MoPPi v.1.1.3.8 (converts binarized .pgn files into a single .xml file); and MultiCompare v.0.9.0 (extracts CMP data from .xml files). A flow chart for TIS image analysis is shown in Supplementary Fig. S1.