Annu Int Conf IEEE Eng Med Biol Soc. 2021 Nov;2021:4844-4850. doi: 10.1109/EMBC46164.2021.9630858.
Microinjection is a widely used technique employed by biologists with applications in transgenesis, cryopreservation, mutagenesis, labeling/dye injection and in-vitro fertilization. However, microinjection is an extremely laborious manual procedure, which makes it a critical bottleneck in the field and thus ripe for automation. Here, we present a computer-guided robot that automates the targeted microinjection of Drosophila melanogaster and zebrafish (Danio rerio) embryos, two important model organisms in biological research. The robot uses a series of cameras to image an agar plate containing embryos at multiple magnifications and perspectives. This imaging is combined with machine learning and computer vision algorithms to pinpoint a location on the embryo for targeted microinjection with microscale precision. We demonstrate the utility of this microinjection robot to successfully microinject Drosophila melanogaster and zebrafish embryos. Results obtained indicate that the robotic microinjection approach can significantly increase the throughput of microinjection as compared to manual microinjection while maintaining survival rates comparable to human operators. In the future, this robotic platform can be used to perform high throughput microinjection experiments and can be extended to automatically microinject a host of organisms such as roundworms (Caenorhabditis elegans), mosquito (Culicidae) embryos, sea urchins (Echinoidea) and frog (Xenopus) oocytes.
显微注射是一种广泛应用于生物学的技术,在转基因、冷冻保存、诱变、标记/染料注射和体外受精等方面都有应用。然而,显微注射是一项极其繁琐的手动操作,这使其成为该领域的一个关键瓶颈,因此非常适合自动化。在这里,我们展示了一种计算机引导的机器人,它可以自动对果蝇(Drosophila melanogaster)和斑马鱼(Danio rerio)胚胎进行靶向显微注射,这两种都是生物学研究中的重要模式生物。机器人使用一系列摄像机以多种放大倍数和视角对含有胚胎的琼脂板进行成像。这种成像与机器学习和计算机视觉算法相结合,以精确定位胚胎上的目标位置,实现微尺度的精确显微注射。我们展示了这种显微注射机器人在成功对果蝇和斑马鱼胚胎进行显微注射方面的实用性。结果表明,与手动显微注射相比,机器人显微注射方法可以显著提高显微注射的通量,同时保持与人工操作人员相当的存活率。在未来,这种机器人平台可以用于进行高通量的显微注射实验,并可以扩展到自动对包括线虫(Caenorhabditis elegans)、蚊子(Culicidae)胚胎、海胆(Echinoidea)和青蛙(Xenopus)卵母细胞在内的一系列生物体进行显微注射。