Amr Abouelezz

Abstract Microscopy underpins modern biology, but acquiring high-quality data from complex experiments at scale remains challenging. Smart microscopy can standardize and automate data collection, but each experiment-specific solution requires coding expertise and development time.

Individual cells were marked, and ITGB5 (or αvβ5) and p-Pax channels were transformed into binary masks using the Robust Automatic Threshold Selection function. The Robust Automatic Threshold Selection parameters used for segmentation (“noise,” “lambda,” and “min”) were defined by visual inspection for each experiment. The mask of the p-Pax channel was then subtracted from the mask of the ITGB5 (or αvβ5) channel using the “image calculation” function.