Continuous long-term detection of live cell surface markers by ‘in culture’ antibody staining

Long-term observation of cell behavior at the single cell level is a crucial prerequisite for understanding molecular cell fate control.This is of particular importance when analyzing highly dynamic and heterogeneous developmental and stem cell systems.The required observations are possible by timelapse microscopy, but reliable identi cation of speci c cell types and maturation stages usually requires detection of molecular marker expression.However, only few of the necessary cell or animal lines expressing genetically encoded uorescent markers exist, and the generation of novel reporter lines is laborious and slow.Here, we describe long-term antibody staining in living cell cultures as an easy, robust, exible and fast way of continuous real-time marker detection on living cells.This approach greatly simpli es the live identi cation of speci c cell types in and dynamic culture systems.