Imaging of individual live cells over time is essential for studying dynamic cellular processes within the heterogeneous cellular population. There is growing interest in using imaging technology to decipher the signaling pathway governing the stimulation of CD4 positive T cells by antigen-presenting cells in order to better understand the associated adaptive immune response. However, there are several technical challenges when conducting time-lapse live cell imaging to study the activation and the differentiation of T cells. First, primary T cells are non-adherent and become highly mobile upon activation. Second, T cell differentiation processes are relatively slow which could take over 72 hours. At last, ability to introduce different reagents as well as a second population of cells to monitor the cellular responses in real time is almost impossible with conventional cell culture substrates. Here we present a microfluidic single cell trapping array platform for dynamic T cell imaging. The platform allows the capture and growth of non-adherent immune cells, long-term monitoring and tracking of cells, separate introduction of antigen presenting cells as well as concurrent imaging with automatic liquid exchange. The protocol to execute each function was developed with a sample experiment demonstrated to screen for cytosolic calcium transients at single cell resolution.
- Copyright © 2013 by The American Association of Immunologists, Inc.