Integrated Biochip-Electronic System with Single-Atom Nanozyme for in Vivo Analysis of Nitric Oxide

Nitric oxide (NO) exhibits a crucial role in various versatile and distinct physiological functions. Hence, its real-time sensing is highly important. Herein, we developed an integrated nanoelectronic system comprising a cobalt single-atom nanozyme (Co-SAE) chip array sensor and an electronic signal processing module (IND_Co-SAE) for both in vitro and in vivo multichannel qualifying of NO in normal and tumor-bearing mice. The high atomic utilization and catalytic activity of Co-SAE endowed an ultrawide linear range for NO varying from 36 to 4.1 × 10⁵ nM with a low detection limit of 12 nM. Combining in situ attenuated total reflectance surface enhanced infrared spectroscopy (ATR-SEIRAS) measurements and density function calculation revealed the activating mechanism of Co-SAE toward NO. The NO adsorption on an active Co atom forms *NO, followed by the reaction between *NO and OH^-, which could help design relevant nanozymes. Further, we investigated the NO-producing behaviors of various organs of both normal and tumor-bearing mice using the proposed device. We also evaluated the NO yield produced by the wounded mouse using the designed device and found it to be approximately 15 times that of the normal mouse. This study bridges the technical gap between a biosensor and an integrated system for molecular analysis in vitro and in vivo. The as-fabricated integrated wireless nanoelectronic system with multiple test channels significantly improved the detection efficiency, which can be widely used in designing other portable sensing devices with multiplexed analysis capability.