The ability to culture cells in vitro has revolutionized hypothesis testing in basic cell and molecular biology research and has become a standard methodology in drug screening and toxicology assays. However, the traditional cell culture methodology - consisting essentially of the immersion of a large population of cells in a homogeneous fluid medium - has become increasingly limiting, both from a fundamental point of view (cells in vivo are surrounded by complex spatiotemporal microenvironments) and from a practical perspective (scaling up the number of fluid handling steps and cell manipulations for high-throughput studies in vitro is prohibitively expensive). The recent advances by our laboratory to address both limitations will be presented, including a microfluidic long-term cell culture platform that features cellular micropatterns and focal delivery of soluble factors to single cells. We are also developing elastomeric sensors and actuators for single-cell probing and manipulation by inexperienced users. These inexpensive technologies allow us to test novel hypotheses concerning neuromuscular development, chemotaxis, and neuronal axon guidance.