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

微型生物任务的自动化和控制：单细胞手术

• 批准号: RGPIN-2018-04814
• 负责人: Mills, James
• 金额: $ 9.25万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=759674
• 关键词: Automation; Control; Micro; scale; Biological

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 ProgramThis 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.

在生物科学领域，过去十年人们对单细胞生物学的兴趣日益浓厚，同时使这项工作成为可能，对开发进行单细胞手术的工具的工程研究的兴趣也日益浓厚。这些单细胞手术过程包括：（i）去除（活检）细胞器，例如细胞。细胞核、线粒体，用于细胞器特性表征，(ii) 注射化学标记物进行细胞内传感，(iii) 将 RNA、DNA 转移到细胞中以创建转基因生物，(iv) 对人类干细胞治疗中干细胞系的细胞成分进行活检，以及 (v) 卵裂球或滋养外胚层活检（人类胚胎细胞）植入前遗传诊断 (PGD) 在孩子发育之前检测遗传疾病。加拿大和世界各地的研究实验室和体外受精 (IVF) 诊所对此类流程的需求不断增加。目前，这些任务是由高技能技术人员使用显微镜和微量移液器完成的。使用明视野显微镜，在显微镜下细胞显得透明。由于景深非常窄（小于几微米），只有细胞的非常薄的切片处于焦点。因此，图像平面上方或下方的感兴趣物体会严重失焦。焦深有限，仅为几微米。再加上操作员的疲劳和潜在的人体污染，对细胞手术的吞吐量和成功率产生了显着的负面影响。通过将自动化制造原理应用于细胞手术任务，并结合实时 3D 图像反馈来控制自动化过程，找到了解决这些问题的方法。消除人类操作员将提高吞吐量和成功率。 为了克服细胞手术中 2D 图像反馈的局限性，我们建议研究和开发一种用于细胞手术的实时 3D 成像系统，该系统将持续提取细胞手术所需的 3D 信息，例如，细胞手术所需的 3D 信息。细胞器质心位置、细胞膜位置和其他量。 研究计划的目标这项拟议的研究旨在研究利用自动化机器人生物操纵系统进行单细胞手术的自动化。 1. 开发机器人驱动的微量移液器自动化细胞手术的自动化策略。2.开发基于 3D 视觉的方法，以促进细胞的定位和后续处理。 3. 处理前将对细胞进行细胞器鉴定。该成像将识别感兴趣的细胞器及其位置。 4. 使用具有视觉反馈的 MEMS 设备开发电磁方法，以两个方向旋转悬浮细胞，用于成像和随后的细胞手术。 5. 开发基于机器人的控制方法，用于使用 3D 图像信息对小细胞进行自动细胞器活检和细胞注射。