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Manipulating single cells with a micropipette is the oldest, yet still a widely used technique. This paper discusses the aspiration of a single cell into a micropipette and positioning the cell accurately to a target position inside the micropipette. Due to the small volume of a single cell (picoliter) and nonlinear dynamics involved, these tasks have high skill requirements and are labor intensive in manual operation that is solely based on trial and error and has high failure rates. We present automated techniques in this paper for achieving these tasks via computer vision microscopy and closed-loop motion control. Computer vision algorithms were developed to detect and track a single cell outside and inside a micropipette for automated single-cell aspiration. A closed-loop robust controller integrating the dynamics of cell motion was designed to accurately and efficiently position the cell to a target position inside the micropipette. The system achieved high success rates of 98% for cell detection and 97% for cell tracking (n = 100). The automated system also demonstrated its capability of aspirating a single cell into a micropipette within 2 s (versus 10 s by highly skilled operators) and accurately positioning the cell inside the micropipette within 8 s (versus 25 s by highly skilled operators).