Due to a traditional optical tweezer microscope has some drawbacks of a bulky shape and a difficulty to operate; therefore, in the study, the researcher utilized two optical pickups and a webcam to build up a simpler optical tweezer system. The system can not only identify the focus point along the axis but control voice coil motor to focus the sample as well. Besides, the system can identify the sample and the laser spot by image processing at the horizontal level to move the sample cell and automatically capture the micro/nano-scale sample.