Herein, a 60-electrode array is fabricated down the length of a microchamber for analysis of a microphysiological system. The electrode array is fabricated by standard photolithographic, metallization, and etching techniques. Permutations of 2-wire impedance measurements (10 Hz to 1 MHz) are made along the length of the microchannel using a multiplexer, Gamry potentiostat, and custom Labview code. An impedance "heat map" is created via custom algorithms. Spatial resolution and mapping capabilities are exhibited using conductive NaCl solutions and 2D cell culture.