Cell migration is an essential characteristic of both physiological and pathological processes within the human body. In order to study the complex process of cell migration different in vitro model systems have been developed in the past. The challenge for all these assays is to provide the cells a substrate that mimics particular properties of the extracellular matrix while a high control over experimental parameters and monitoring is desired. However, migration assays commonly used in cell biology and medical research are rather limited in the control over the architecture of the provided matrix on or through which the cells move or by the lack of adequate imaging devices to monitor cell dynamics. To overcome some limitations of conventional migration assays, it was the aim of this work to develop two different methods and employ them in order to quantify migrative behavior of cells under precisely controlled in vitro conditions.