Peripheral blood is the most readily available resource for stroke patient prognosis, but there is a lack of methods to detect dynamic changes of neutrophils in peripheral blood that can be used in the clinic. Herein, we developed a procedure to characterize dynamic changes of neutrophils based on their electrical properties in rats after transient middle cerebral artery occlusion (MCAO). We characterized the specific membrane capacitance (Csm) and cytoplasmic resistance (σcyto) of approximately 27,600 neutrophils from MCAO rats 24 h after ischemia/reperfusion. We found that the Csm and σcyto of neutrophils in the MCAO group were significantly higher compared to the sham group. Furthermore, we observed a monotonically upward shift in neutrophil Csm in the MCAO group during the four 5-minute test cycles. Our findings suggest that the dynamic changes of cellular electrical properties could reflect neutrophil activity and serve as a prognostic indicator for ischemic stroke in the clinical setting.