Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, M5S, Canada.
Biomed Microdevices. 2019 Jun 15;21(3):52. doi: 10.1007/s10544-019-0391-z.
Micromanipulation is the precise in vitro handling and study of individual biological cells, where the smallest error can be disastrous. One such example is the extraction of cellular material from multicellular organisms, such as cells from early stage embryos. In this paper, we propose automation methods for the extraction and retrieval of individual cells from a multicellular organism in vitro using the displacement method. Computer-controlled syringe pumps and micromanipulators combined with custom computer vision algorithms are used for automated cell extraction and retrieval. Automation feasibility is demonstrated through automated controlled extraction of one or two blastomeres from cleavage-stage embryos. Preliminary proof of concept blastomere extraction experiments involving mouse embryos obtained success rates ranging from 72% to 88% for the different extraction tasks: displacement, detection, and retrieval. These automated blastomere extraction experiments demonstrate that automated cell extraction is indeed feasible, but the process may still be improved. To the best of these authors' knowledge, this paper is the first to report the automation of single cell extraction from multicellular organisms using the displacement method, and especially for automated blastomere extraction from cleavage-stage embryos. These methods provide a set of tools for moving towards fully automated single cell surgery procedures.
微操作是指对单个生物细胞进行精确的体外操作和研究，任何微小的误差都可能造成灾难性的后果。例如，从多细胞生物中提取细胞物质，如早期胚胎中的细胞。在本文中，我们提出了一种使用位移法从体外的多细胞生物中提取和回收单个细胞的自动化方法。计算机控制的注射器泵和微操作器与定制的计算机视觉算法相结合，用于自动细胞提取和回收。通过自动控制从分裂期胚胎中提取一个或两个卵裂球，证明了自动化的可行性。涉及小鼠胚胎的初步概念验证卵裂球提取实验，在不同的提取任务（位移、检测和回收）中成功率从 72%到 88%不等。这些自动卵裂球提取实验表明，自动细胞提取确实是可行的，但该过程仍有待改进。据作者所知，本文首次报道了使用位移法从多细胞生物中自动提取单个细胞，特别是从分裂期胚胎中自动提取卵裂球。这些方法为实现全自动单细胞手术程序提供了一套工具。
