High-resolution and high-speed 3D tracking of microrobots using a fluorescent light field microscope

Background: Imaging and tracking are crucial for microrobots which navigate through complex 3D environments. Fluorescent imaging (FI) by microscope offers a high-resolution and high-sensitive imaging method to study the property of microrobots. However, conventional microscope suffers from shallow depth of field (DOF) and lacks 3D imaging capability. Methods: We proposed a high-resolution and high-speed 3D tracking method for microrobots based on a fluorescent light field microscope (FLFM). We designed the FLFM system according to the size of a representative helical microrobot (150 μm body length, 50 μm screw diameter), and studied the system’s performance. We also proposed a 3D tracking algorithm for microrobots using digital refocusing. Results: We validated the method by simulations and built an FLFM system to perform the tracking experiments of microrobots with representative size. Our 3D tracking method achieves a 30 fps data acquisition rate, 10 μm lateral resolution and approximately 40 μm axial resolution over a volume of 1,200×1,200×326 μm³. Results indicate that the accuracy of the method can reach about 9 μm. Conclusions: Compared with the FI by a conventional microscope, the FLFM-based method gains wider DOF and 3D imaging capability with a single-shot image. The tracking method succeeds in providing the trajectory of the microrobot with a good lateral resolution.