The tracking of intracellular organelles and vesicles is becoming increasingly important for understanding cellular dynamics. Originally, the development of tracking algorithms was mainly pursued in other fields, e.g. aerospace/military/street surveillance. However, most of this algorithm is not directly applicable to live cell microscopy data. Here we describe the algorithms that have been successfully applied to object detection and tracking specifically in vivo and in vitro motility assays. The characteristics advantages and disadvantages of the different approaches are compared.