Abstract: In order to track the ultrasonic imaging plane using a 3D spatial tracking device in application such as ultrasound guided interventional operation, a transform between ultrasound probe and the ultrasound image has to be determined by calibration. Based on the N-phantom, we use a phantom that 8 points is known and NDI optical positioning system for calibration to do this. The calibration process divides into three steps:firstly getting 8 bright spots orderly in the ultrasound image when the phantom is scanned by the ultrasound probe with a receiver sensor; secondly using the NDI optical positioning system get the transform between receiver and transmitter coordinate, the transmitter and phantom coordinate; lastly realizing the calibration by extracting feature points of the image bright spots and deciding the position relation matrix according to least square method. It is proved that the calibration method is convenient to use, and the calibration precision is not only meet the demand of ultrasound guide and exceed to conventional approach and reach to 2.72 mm,laying a good foundation for further 3D reconstruction.