Holographic optical tweezers (HOT) technology is a new experimental technology. It perfectly combines holographic and optical tweezers and becomes an indispensable scientific research tool in the interdisciplinary field. This article discusses the transformation of the holographic optical tweezers experiment into an optical comprehensive teaching experiment. Through this comprehensive teaching experiment, students can understand the design and calibration technology of holographic optical tweezers, master the structure, principle and operation method of the holographic optical tweezers system, and deepen their understanding of the mechanical effects of light and holographic principles. This experiment helps to improve students comprehensive practical ability, stimulate students interest in learning, cultivate students innovative consciousness.