Cell extraction automation in single cell surgery using the displacement method.

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.