This paper aimed at investigating the properties of cultured cardiomyocytes using microcirculatory and molecular technology to culture cardiomyocytes from different parts of the neonate Wistar rat's heart and record their spontaneous pulsation under time-lapse video microscopy, then analyze their activity and inspect their survival rate and apoptotic rate under natural and nitric oxide conditions. The pulsation frequency in cardiomtocytes of different parts in heart are: 78.5+/-11.0 beats/min in the atrium, 88.4+/-6.3 beats/min in the left ventricle, 90.3+/-7.9 beats/min in the right ventricle and 115.3 +/-11.4 beats/min in the cardiac apex, respectively, with an average frequency of 81.3 beats/min. Different concentrations of nitric oxide showed no effect on the frequency of cardiomyocyte pulsation. The survival rates of the above cardiomyocytes are 96.0%, 95.0%, 95.0%, and 95.3% respectively and 95.0% for the whole heart. The apoptotic rates are 1.3%, 1.1%, 4.8%, and 1.8% respectively and 5.1% for the whole heart. Different concentrations of nitric oxide had no effect on these results. Our study showed that cultured myocardial cells from different parts of the heart displayed various pulsation frequencies, and the frequency of the cardiac apex is the highest while the atrium is lowest. We also found that there is no statistically significant difference in the survival rates and apoptotic rates of different parts of the heart, and that nitric oxide has no effect on the beating frequency, survival rates or apoptotic rates of the cardiomyocytes in vitro.