Department of Biomedical Engineering, University of Minnesota, Twin Cities, MN, USA.
Department of Biomedical Engineering, Duke University, Durham, NC, USA.
EMBO Rep. 2019 Oct 4;20(10):e47880. doi: 10.15252/embr.201947880. Epub 2019 Aug 30.
Microinjection into single cells in brain tissue is a powerful technique to study and manipulate neural stem cells. However, such microinjection requires expertise and is a low-throughput process. We developed the "Autoinjector", a robot that utilizes images from a microscope to guide a microinjection needle into tissue to deliver femtoliter volumes of liquids into single cells. The Autoinjector enables microinjection of hundreds of cells within a single organotypic slice, resulting in an overall yield that is an order of magnitude greater than manual microinjection. The Autoinjector successfully targets both apical progenitors (APs) and newborn neurons in the embryonic mouse and human fetal telencephalon. We used the Autoinjector to systematically study gap-junctional communication between neural progenitors in the embryonic mouse telencephalon and found that apical contact is a characteristic feature of the cells that are part of a gap junction-coupled cluster. The throughput and versatility of the Autoinjector will render microinjection an accessible high-performance single-cell manipulation technique and will provide a powerful new platform for performing single-cell analyses in tissue for bioengineering and biophysics applications.
将微量注射技术应用于脑组织中的单细胞是研究和操纵神经干细胞的一种强大技术。然而，这种微量注射需要专业知识，并且是一个低通量的过程。我们开发了“自动注射器”，这是一种机器人，它利用显微镜图像引导微注射针进入组织，将皮升体积的液体注入单个细胞。AutoInjector 能够在单个器官型切片中注射数百个细胞，其总体产量比手动微量注射高出一个数量级。AutoInjector 能够成功地将胚胎小鼠和人胎大脑中的顶侧祖细胞 (APs) 和新生神经元作为目标。我们使用 Autoinjector 系统地研究了胚胎小鼠大脑皮层中神经祖细胞之间的缝隙连接通讯，发现顶端接触是缝隙连接偶联簇的一部分细胞的特征。AutoInjector 的高通量和多功能性将使微量注射成为一种易于使用的高性能单细胞操作技术，并为在组织中进行生物工程和生物物理应用的单细胞分析提供一个强大的新平台。
