AIM: To grope the mothod used to isolate adult mice cardiac myocytes and observe the electrophysiological characteristics of the myocytes. METHODS: Enzyme digestion method was used to isolate single cells and whole-cell patch clamp recording technique was used to record Ca 2+ and K + currents. RESULTS: Rod-shaped, clear-striped cardiac myocytes with normal electrophysiological characteristics were gotten. After exposed to normal calcium concentration solution, 50%~60% calcium-resistant myocytes were left. Giga-ohm seal can be easily achieved and Ca 2+ and K + currents were successfully recorded. CONCLUSION: It is an economic, convenient and reliable method to isolate adult mice cardiac myocytes with normal electrophysiological characteristics.