Single-cell information extraction and viability analysis using automated microscopy

We present the use of an automated microscope routine for long-term single cell viability analysis. Murine macrophage cells were monitored for more than 10 hours at physiological conditions. The information of each cell was extracted from time-lapse raw fluorescence and bright-field images to study single cell dynamic behaviors. Two methods were applied to analyze single cell viability: the popular method using live/dead fluorescent dye, and a new morphology-based, dye-free method that estimates viability with optical appearance. Both methods yielded similar death event estimation, indicating the new morphology-based method can be an alternative when using live/dead fluorescent dye is difficult or not allowed