Genetically encoded fluorescent sensors continue to be developed and improved. If they could be expressed across multiple cortical areas in non-human primates, it would be possible to measure a variety of spatiotemporal dynamics of primate-specific cortical activity. Here, we develop an Automated Robotic Virus injection System (ARViS) for broad expression of a biosensor. ARViS consists of two technologies: image recognition of vasculature structures on the cortical surface to determine multiple injection sites without hitting them, and robotic control of micropipette insertion perpendicular to the cortical surface with 50 μm precision. In mouse cortex, ARViS sequentially injected virus solution into 100 sites over a duration of 100 min with a bleeding probability of only 0.1% per site. Furthermore, ARViS successfully achieved 266-site injections over the frontoparietal cortex of a female common marmoset. We demonstrate one-photon and two-photon calcium imaging in the marmoset frontoparietal cortex, illustrating the effective expression of biosensors delivered by ARViS.

基因编码荧光传感器不断得到开发和改进。如果它们可以在非人类灵长类动物的多个皮质区域表达，那么就有可能测量灵长类动物特异性皮质活动的各种时空动态。在这里，我们开发了一种自动机器人病毒注射系统（ARViS），用于生物传感器的广泛表达。 ARViS 由两项技术组成：皮质表面脉管系统结构的图像识别，以确定多个注射部位而无需撞击它们，以及机器人控制垂直于皮质表面插入的微量移液器，精度为 50 μm。在小鼠皮质中，ARViS 在 100 分钟内将病毒溶液顺序注射到 100 个部位，每个部位的出血概率仅为 0.1%。此外，ARViS 成功地对雌性普通狨猴的额顶皮层进行了 266 个部位的注射。我们在狨猴额顶叶皮质中展示了单光子和双光子钙成像，说明了 ARViS 传递的生物传感器的有效表达。