Objective To establish a stable rat model of middle cerebral artery occlusion (MCAO) assisted by laser Doppler flowmetry. Methods Sixteen rats were randomly divided into two groups: (1) experimental group, n=8; (2) control group, n=8. In the experimental group, a sudden decrease of regional cerebral blood flow from baseline (before ischemia) to approximately 30% of baseline confirmed the correct placement of nylon thread. In the control group, 1.8 cm nylon thread insertion was performed without laser Doppler flowmetry. The modified neurological severity scores (mNSS) were tested before and 24 h after MCAO. All rats were killed at 24 h after MCAO. The brains were removed and dissected into 6 pieces of 2 mm coronal sections. The fresh brain slices were stained with 2% 2,3,5-triphenyl tetrazolium chloride and the relative infarct volume was measured.Results In the experimental group, 8 rats showed typical symptom of hemiplegy with stable mNSS (between 10 and 13) and infarct volume (37.5±3.9%). However, only 5 rats in the control group showed stable mNSS and infarct volume. The success rate of the model establishment was 100% in the experimental group compared with 62.5% in the control group (P<0.05). Conclusion The application of laser Doppler flowmetry significantly increases the establishment success rate of stable MCAO models in rats.