Department of Biomedical Engineering, University of Wisconsin-Madison.
Department of Biomedical Engineering, University of Wisconsin-Madison; University of Wisconsin Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health;
J Vis Exp. 2023 Aug 4(198). doi: 10.3791/65686.
Optogenetics offers precise control over cellular behavior by utilizing genetically encoded light-sensitive proteins. However, optimizing these systems to achieve the desired functionality often requires multiple design-build-test cycles, which can be time-consuming and labor-intensive. To address this challenge, we have developed Lustro, a platform that combines light stimulation with laboratory automation, enabling efficient high-throughput screening and characterization of optogenetic systems. Lustro utilizes an automation workstation equipped with an illumination device, a shaking device, and a plate reader. By employing a robotic arm, Lustro automates the movement of a microwell plate between these devices, allowing for the stimulation of optogenetic strains and the measurement of their response. This protocol provides a step-by-step guide on using Lustro to characterize optogenetic systems for gene expression control in the budding yeast Saccharomyces cerevisiae. The protocol covers the setup of Lustro's components, including the integration of the illumination device with the automation workstation. It also provides detailed instructions for programming the illumination device, plate reader, and robot, ensuring smooth operation and data acquisition throughout the experimental process.
光遗传学通过利用基因编码的光敏感蛋白来提供对细胞行为的精确控制。然而，优化这些系统以实现所需的功能通常需要多次设计-构建-测试循环，这可能既耗时又费力。为了解决这个挑战，我们开发了 Lustro，这是一个将光刺激与实验室自动化相结合的平台，能够实现高效的高通量筛选和光遗传学系统的表征。Lustro 利用配备有照明设备、摇动设备和板读数器的自动化工作站。通过使用机械臂，Lustro 自动在这些设备之间移动微孔板，从而可以刺激光遗传学菌株并测量它们的响应。本协议提供了使用 Lustro 对 budding yeast Saccharomyces cerevisiae 中的基因表达控制进行光遗传学系统表征的逐步指南。该协议涵盖了 Lustro 组件的设置，包括将照明设备与自动化工作站集成。它还提供了有关如何对照明设备、板读数器和机器人进行编程的详细说明，以确保整个实验过程中的平稳运行和数据采集。
