This tutorial outline a basic image analysis workflow for Live Cell Painting with ChromaLIVE™ in CellProfiler™. This guide details a simple workflow from preprocessing (uploading images, extracting metadata) to segmentation (identifying nuclei and cells) and feature extraction, aiming to provide multiparametric information crucial for precise phenotypic profiling.

It explains how ChromaLIVE™, our non-toxic dye, enables kinetic measurements of phenotypic changes in live cells at high-throughput due to its biocompatibility and is highlighted as highly compatible with live-cell imaging. The guide further explores both unsupervised (dimensionality reduction, clustering with StratoMineR™) and supervised (classification with CellProfiler Analyst™) data analysis approaches to process extracted features. This comprehensive approach supports Live Cell Painting assays and improves efficiency in compound profiling.