Simultaneous Non-Invasive Electrocardiogram and Respiration Rate Recordings in Head-Fixed Awake Mice.

Autonomic regulation of heart and respiratory rates is essential for understanding brain-body interactions in health and disease. Preclinical cardiovascular recordings are often performed under anesthesia or via telemetry, both of which introduce physiological confounds such as stress or impaired recovery due to the need for acute or chronic implantation of sensors. Here, we present a minimally invasive protocol for simultaneous acquisition of high-quality electrocardiography and respiratory signals in awake mice. Using an in-house-modified physiological monitor in awake, head-fixed mice that were briefly habituated to experimental conditions, we ultimately enable stable, long-term physiological recordings alongside in vivo microscopy. This protocol provides a robust, low-stress method for acquiring physiological signals, enabling the simultaneous study of cardiovascular-cerebral dynamics in awake head-fixed mice, thereby enhancing the translational relevance of preclinical measurements. Key features • Minimally invasive electrocardiogram and respiration rate acquisition in awake, head-fixed mice suitable for long-term physiological recordings. • Custom-built setup integrates physiological monitoring with in vivo imaging without surgical implantation or telemetry. • Rapid habituation protocol ensures low-stress conditions and high-quality signal acquisition in conscious mice. • Enables correlation of cardiovascular dynamics with brain activity and cerebrospinal fluid flow in translational neuroscience studies.