We have developed a methodology for electrohydrodynamic printing of a nitric oxide (NO) sensor. The primary features of this amperometric methodology include the printing of carbon and metallic-based inks, the conversion of Ag to Ag:AgCl as a reference electrode, and the deposition of a nitric oxide selective layer on the working electrode, followed by an encapsulation to define the working area. The efficacy and enhancements of multiple printed layers has been examined. Multiple working electrode chemistries and combinations are presented, including the use of silver, graphene, NiTsPc, and gold in various combinations. Selectivity for NO was accomplished via a NafionO layer on the working electrode, effectively blocking interference from NO2- and NO3- at relatively high concentrations. The resultant flexible, thin film nitric oxide sensor is sensitive enough to monitor macrophage depolarization events without the need for additional micropipette or FRET-based electrochemical sensors. We demonstrate a valuable research tool that will have a high impact on the emerging field of synthetic biology by providing the capability to interface-engineered cellular sensors to microelectronics. [GRAPHICS] .