Precise Automated Intracellular Delivery Using a Robotic Cell Microscope System with Three-Dimensional Image Reconstruction Information

Precise intracellular delivery is essential for many biomedical applications, such as genome-editing and intracellular mechanism investigation. This article presents the development of an automated intracellular delivery approach using a robot-aided microscope system with 3-D reconstruction information. A series of optical section images is sampled by objective lens movement and restored on the computer. After several processes, including deconvolution, segmentation, and 3-D reconstruction of sampled images, the optimal delivery position can be acquired via geometry analysis based on the proposed volume-based model. Guided by this 3-D information, a robust controller that can compensate for external disturbances is utilized to manipulate micropipettes to deliver exogenous materials into single cells. Reconstruction accuracy experiments are performed on the cell nucleus, and the results are compared with a confocal fluorescent microscope. Intracellular nucleus delivery experiments are further conducted by using the proposed robotic microscope system. Compared with the widely adopted methods without 3-D information, the proposed approach demonstrates substantial improvement.