Nanomaterials show great potential for applications in biosensing due to their unique physical, chemical, and biological properties. However, the single-modal signal sensing mechanism greatly limits the development of single-modal nanoprobes and their related sensors. Multi-modal nanoprobes can realize the output of fluorescence, colorimetric, electrochemical, and magnetic signals through composite nanomaterials, which can effectively compensate for the defects of single-modal nanoprobes. Following the multi-modal nanoprobes, multi-modal biosensors break through the performance limitation of the current single-modal signal and realize multi-modal signal reading. Herein, the current status and classification of multi-modal nanoprobes are provided. Moreover, the multi-modal signal sensing mechanisms and the working principle of multi-modal biosensing platforms are discussed in detail. We also focus on the applications in pharmaceutical detection, food and environmental fields. Finally, we highlight this field's challenges and development prospects to create potential enlightenment.