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However, the manual operation needs a skilled operator and faces problems such as low throughput and poor reproducibility. This paper proposes an automated nanopipette-based microoperation system for cell detection, three-dimensional nonovershoot positioning of the nanopipette tip in proximity to the cell of interest, cell approaching and proximity detection between nanopipette tip and cell surface, and cell penetration and detection of the intracellular reactive oxygen species (ROS). A robust focus algorithm based on the number of cell contours was proposed for adherent cells, which have sharp peaks while retaining unimodality. The automated detection of adherent cells was evaluated on human umbilical cord vein endothelial cells (HUVEC) and NIH\/3T3 cells, which provided an average of 95.65% true-positive rate (TPR) and 7.59% false-positive rate (FPR) for in-plane cell detection. The three-dimensional nonovershoot tip positioning of the nanopipette was achieved by template matching and evaluated under the interference of cells. Ion current feedback was employed for the proximity detection between the nanopipette tip and cell surface. Finally, cell penetration and electrochemical detection of ROS were demonstrated on human breast cancer cells and zebrafish embryo cells. This work provides a systematic approach for automated intracellular sensing for adherent cells, laying a solid foundation for high-throughput detection, diagnosis, and classification of different forms of biochemical reactions within single cells.<\/jats:p>","DOI":"10.34133\/2022\/9763420","type":"journal-article","created":{"date-parts":[[2022,9,2]],"date-time":"2022-09-02T23:45:10Z","timestamp":1662162310000},"source":"Crossref","is-referenced-by-count":38,"title":["Robotic Intracellular Electrochemical Sensing for Adherent Cells"],"prefix":"10.34133","volume":"2022","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8315-7251","authenticated-orcid":false,"given":"Weikang","family":"Hu","sequence":"first","affiliation":[{"name":"Shenzhen Key Laboratory of Biomimetic Robotics and Intelligent Systems, Department of Mechanical and Energy Engineering, Southern University of Science and Technology, Shenzhen, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanmei","family":"Ma","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Biomimetic Robotics and Intelligent Systems, Department of Mechanical and Energy Engineering, Southern University of Science and Technology, Shenzhen, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhen","family":"Zhan","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Biomimetic Robotics and Intelligent Systems, Department of Mechanical and Energy Engineering, Southern University of Science and Technology, Shenzhen, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danish","family":"Hussain","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Biomimetic Robotics and Intelligent Systems, Department of Mechanical and Energy Engineering, Southern University of Science and Technology, Shenzhen, China"},{"name":"Department of Mechatronics Engineering, National University of Sciences and Technology, Islamabad, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8627-2335","authenticated-orcid":false,"given":"Chengzhi","family":"Hu","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Biomimetic Robotics and Intelligent Systems, Department of Mechanical and Energy Engineering, Southern University of Science and Technology, Shenzhen, China"},{"name":"Guangdong Provincial Key Laboratory of Human-Augmentation and Rehabilitation Robotics in Universities, Southern University of Science and Technology, Shenzhen, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2796"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-020-0230-3"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1003428107"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2010.08.040"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aat0271"},{"key":"e_1_3_4_6_2","doi-asserted-by":"crossref","DOI":"10.34133\/2021\/1532103","article-title":"Challenges and possibilities of cell-based tissue-engineered vascular grafts","volume":"2021","author":"Saito J.","year":"2021","unstructured":"J. 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