Cornell E, Fisher W W, Nordmeyer R, Yegian D, Dong M, Biggin M D, Celniker S E, Jin J
Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Rev Sci Instrum. 2008 Jan;79(1):013705. doi: 10.1063/1.2827516.
To decipher and manipulate the 14 000 identified Drosophila genes, there is a need to inject a large number of embryos with transgenes. We have developed an automated instrument for high throughput injection of Drosophila embryos. It was built on an inverted microscope, equipped with a motorized xy stage, autofocus, a charge coupled device camera, and an injection needle mounted on a high speed vertical stage. A novel, micromachined embryo alignment device was developed to facilitate the arrangement of a large number of eggs. The control system included intelligent and dynamic imaging and analysis software and an embryo injection algorithm imitating a human operator. Once the injection needle and embryo slide are loaded, the software automatically images and characterizes each embryo and subsequently injects DNA into all suitable embryos. The ability to program needle flushing and monitor needle status after each injection ensures reliable delivery of biomaterials. Using this instrument, we performed a set of transformation injection experiments. The robot achieved injection speeds and transformation efficiencies comparable to those of a skilled human injector. Because it can be programed to allow injection at various locations in the embryo, such as the anterior pole or along the dorsal or ventral axes, this system is also suitable for injection of general biochemicals, including drugs and RNAi.
为了解码和操控已识别出的14000个果蝇基因,需要向大量胚胎中注射转基因。我们开发了一种用于果蝇胚胎高通量注射的自动化仪器。它基于一台倒置显微镜构建,配备了电动xy载物台、自动聚焦装置、电荷耦合器件相机以及安装在高速垂直载物台上的注射针。开发了一种新型的微机械加工胚胎对齐装置,以方便大量卵的排列。控制系统包括智能动态成像与分析软件以及模仿人类操作员的胚胎注射算法。一旦装载了注射针和胚胎载玻片,软件会自动对每个胚胎进行成像和特征描述,随后将DNA注入所有合适的胚胎。每次注射后对针进行冲洗编程和监测针状态的能力确保了生物材料的可靠递送。使用该仪器,我们进行了一系列转化注射实验。该机器人实现的注射速度和转化效率与熟练的人类注射者相当。由于它可以编程以允许在胚胎的不同位置进行注射,如前极或沿背轴或腹轴,该系统也适用于注射包括药物和RNA干扰在内的一般生化物质。