The invention provides a multifunctional microvesicle which integrates ultrasonic/fluorescent dual-mode tomography and gene therapy/photodynamic therapy. According to embodiment results, the multifunctional microvesicle has excellent gene transfection capacity and singlet oxygen generation capacity, has ultrasonic developing enhancement property and fluorescent imaging capacity, and can effectively inhibit growth of tumor cells. The invention further provides a preparation method of the multifunctional microvesicle. The method is simple and convenient to operate and easy to implement. The invention further provides an application of the multifunctional microvesicle, wherein the multifunctional microvesicle can be used for preparing cancer diagnosis or therapy reagents.