3-D image reconstruction of biological organelles with a robot-aided microscopy system for intracellular surgery

Intracellular surgery suffers from a low success rate due to the lack of three-dimensional (3-D) position feedback of selected organelles. In this study, we developed a novel robot-aided microscopy system and 3-D reconstruction algorithm to conduct intracellular surgery with 3-D information. A series of optical sections along the vertical direction was obtained by microscopy lens movement. A 3-D reconstruction model of the specimen was realized after several deconvolution, segmentation, and reconstruction processes. Simulations and experiments were performed to verify the accuracy of the proposed algorithm. Furthermore, 3-D reconstruction position feedback was applied to extract mitochondria via a robot-aided wide-field fluorescence system. The proposed approach facilitates intracellular surgeries, such as organelle biopsy and cell injection.