Objective：To compare the accuracyof interactive closet point (ICP) algorithm, Procrustes analysis (PA) algorithm,andalandmark-independent method to construct the mid-sagittal plane (MSP) of the cone beam computed tomography.To provide theoretical basis for establishing coordinate systemof CBCT images and symmetric analysis. Methods: Ten patients were selected and scanned by CBCT before orthodontic treatment.The scan data was imported into Mimics 10.0 to reconstructthree dimensional skulls.And the MSP of each skull was generated by ICP algorithm, PA algorithm and landmark-independent method. MSP extracted by ICP algorithm or PA algorithm involvedthree steps. First, the 3D skull processing was performed by reverse engineering software geomagic studio 2012 to obtain the mirror skull. Then, the original and its mirror skull was registered separately by ICP algorithm in geomagic studio 2012 and PA algorithm in NX Imageware 11.0. Finally, the registered data were united into new data to calculate the MSP of the originaldata in geomagic studio 2012. The mid-sagittal plane was determined by SELLA (S), nasion (N), basion (Ba) as traditional landmark-dependent methodconducted in software InVivoDental 5.0. The distance from 9 pairs of symmetric anatomical marked points to three sagittal plane were measured and calculated to compare the differences of the absolute value. The one-way ANOVA test was used to analyze the variable differences among the 3 MSPs. The pairwise comparison was performed with LSD method. Results: MSPs calculated by the three methods were available for clinic analysis, which could be concluded from the front view.However, there was significant differences among the distances from the 9 pairs of symmetric anatomical marked points to the MSPs (F=10.932，P=0.001).LSD test showed there was no significant difference between the ICP algorithm and landmark-independent method (P=0.11), while there was significant difference between the PA algorithm and landmark-independent methods (P=0.01) .Conclusion: Mid-sagittal plane of 3D skulls could be generated base on ICP algorithm or PA algorithm. There was no significant difference between the ICP algorithm and landmark-independent method. For the subjects with no evident asymmetry, ICP algorithm is feasible in clinical analysis.