Cells in tissues and organs coordinate their activities by communicating with each other, and this communication is mediated by specialized channels, called gap junctions. Through the gap junction channels, cells synchronize their electrical and metabolic activities, transmit intercellular signals, resulting in regulation of cell growth and differentiation, and maintenance of metabolic homeostasis. This proposal aims to develop a novel microfluidic sensor that is capable of probing intercellular communications via gap junction channels at the single-cell level. The proposed approach utilizes the multi-stream laminar flow characteristics of a microfluidic chip to precisely confine adjacent cells to different environments, thereby enabling a controlled means to study intercellular communications between a single pair of cells in real-time by both fluorescence and/or impedance methods. The sensor would provide an ability to study the specificity of reagents and response of gap junctions to extracellular stimuli. Several research goals will be accomplished in this project, including design and control of the multi-stream flow in a microfluidic channel, on-chip cell culture in the functionalized regions of the sensor, detection of intercellular communication pathways using fluorescence and impedance measurements and their response to extracellular stimuli, and develop sensing strategies for studying water permeability of gap junction channels.
