Multiscale, multi-perspective imaging assisted robotic microinjection of 3D biological structures.
Multiscale, multi-perspective imaging assisted robotic microinjection of 3D biological structures.
复制标题
DOI：
10.1109/embc46164.2021.9630858
复制
发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Kodandaramaiah SB
中科院分区：
文献类型：
--
作者：
Joshi AS;Alegria AD;Auch B;Khosla K;Mendana JB;Liu K;Bischof J;Gohl DM;Kodandaramaiah SB
显微注射是生物学家广泛使用的技术，其应用于转基因、冷冻保存、诱变、标记/染料注射和体外受精。然而，显微注射是一个极其费力的手动过程，这使得它成为该领域的关键瓶颈，因此自动化的时机已经成熟。在这里，我们提出了一个计算机引导的机器人，自动化有针对性的显微注射果蝇和斑马鱼（Danio rerio）胚胎，两个重要的模式生物在生物研究。该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，以确定胚胎上的位置，以便以微尺度精度进行靶向显微注射。我们证明了这种显微注射机器人的效用，成功地显微注射果蝇和斑马鱼胚胎。获得的结果表明，机器人显微注射方法可以显着增加微注射的吞吐量相比，手动显微注射，同时保持与人类操作员相当的生存率。在未来，这个机器人平台可用于进行高通量显微注射实验，并可扩展到自动显微注射一系列生物体，如蛔虫（秀丽隐杆线虫），蚊子（蚊科）胚胎，海胆（海胆）和青蛙（非洲爪蟾）卵母细胞。
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.