Objective To design and construct a rodent awake activity device for brain microdialysis recording and achieve the application and verification of brain microdialysis sampling in mice. Methods The rodent awake activity device for brain microdialysis recording was designed and constructed using relevant materials. Dopamine-related neurotransmitters in the substantia nigra-striatum neural circuit in mice were measured and verified by this device. Microdialysis probes were implanted into the substantia nigra and striatum. Recording and microdialysis sampling were performed for 7 days after operation. Dopamine and its metabolic intermediate DOPAC were analyzed in the substantia nigra and striatum. The microdialysis flow rate was 2 μL/ minute. Samples were collected every 20 minutes for 200 minutes to analyze DA and DOPAC concentration changes over time. Results ( 1) Mouse brain samples showed stable levels of neurotransmitters. (2)Dopamine concentrations were (0. 11 ± 0. 01) nmol / L in the substantia nigra and (0. 22 ± 0. 05) nmol / L in the striatum. DOPAC concentrations were (3. 38 ± 0. 59) nmol / L in the substantia nigra and (20. 35 ± 6. 87) nmol / L in the striatum. No significant differences were found between each sampling period. Conclusions The rodent awake activity device for brain microdialysis recording is practical and stable with good applications in time and space dimensions. Two national patents have been applied for and authorized: Patent for invention ( 202110525342. 5) and Patent for utility models (202121027247. 4).