Automation and Control of Micro-scale Biological Tasks: Single Cell Surgery

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Within the biological sciences, the last decade has witnessed growing interest in single cell biology, and simultaneously enabling this work, increasing interest within engineering research to develop tools to carry out single cell surgery. These single cell surgery processes include: (i) removal (biopsy) of cell organelles e.g. nucleus, mitochondria, for organelle property characterization, (ii) injection of chemical markers for intracellular sensing, (iii) transfer of RNA, DNA into the cell to create transgenic organisms, (iv) biopsy of cell components for stem cell lines in human stem cell therapy and (v) blastomere or trophectoderm biopsy (human embryonic cell) Preimplantation Genetic Diagnosis (PGD) to detect genetic disease prior to the development of a child. Increasing demand for such processes is seen in both research laboratories and in-vitro fertilization (IVF) clinics in Canada and worldwide. Currently, these tasks are carried out by highly skilled technicians, using microscopes, with micro-pipettes. Bright field microscopes are used, under which cells appear transparent. Due to the very narrow depth of field (less than a few microns), only a very thin slice of the cell is in focus. Hence, objects of interest above or below the image plane, are extremely out of focus. This limited depth of focus of only a few µm. coupled with operator fatigue and potential human contamination, contributes to a significant negative impact on throughput and success rate of cell surgery. A solution to these issues is found through the application of automated manufacturing principles to cell surgery tasks, coupled with real-time 3D image feedback to control the automation processes. Elimination of human operators will result in increased throughput and success rates. To overcome the limitations of 2D image feedback in cell surgery, we propose to investigate and develop a real-time 3D imaging system for cell surgery which will continuously extract 3D information essential for cell surgery, e.g. organelle centroid location, cell membrane location, and other quantities. Objectives of the Research Program This proposed research aims to investigate automation of single cell surgery with an automated robotic bio-manipulation system. 1. Develop automation strategies for robotic actuated micro-pipette automated cell surgery. 2. Develop 3D vision based methodologies to facilitate the location and subsequent processing of cells. 3. Cell organelle identification will be carried out on cells prior to processing. This imaging will identify the cell organelle of interest, and its location. 4. Develop electromagnetic approaches using MEMS device with vision feedback, to rotate suspended cells in two direction rotations, for imaging and subsequent cell surgery. 5. Develop robotic based control methods for automated cell organelle biopsy and cell injection for small cells using 3D image information.