A Versatile Automated Platform for Micro-scale Cell Stimulation Experiments

The overall goal of this procedure is to assemble and use a semi-automated microfluidic based platform to cultivate and stimulate small populations of cells and recover cell lysates for molecular analyses. This is accomplished by first establishing microscale cell cultures in fibronectin coated micro capillaries referred to as cell perfusion chambers. The second step is to connect the perfusion chambers to a digital microfluidics or DMF module, which provides a flexible means by which reagents are routed to and from the microscale cell cultures.

Next, the DMF module delivers the stimulus of interest to the perfusion chambers and the cells are incubated with the stimulus for a predetermined time of exposure. Finally, the cells are lysed by chemical means and the lysates are recovered onto the DMF module for molecular analysis. The recovered lysates are then analyzed using quantitative R-T-P-C-R in order to characterize the cell population's transcriptional response to the stimulus.