Clearbot: a system for fully automated, high-throughput tissue clearing and immunostaining

总结 组织清除是一种强大的技术，它使大的生物样品光学透明，并允许3D 宽脑分子解剖的可视化。为了帮助推广这项技术，半透明生物系统公司 开发了Mesoscale Imaging System™，并利用先前的SBIR NIH支持，开发了一个分析软件 用于大脑样本高级分析的软件包，3 TKTM。虽然存在成像和分析的完整解决方案， 对于高劳动强度和易出错的组织清除， 过程在这里，半透明生物系统公司介绍了Clearbot™，这是一种用于样品清洗和 免疫染色，将补充以前的半透明设备，并完成一个管道，可靠的大脑- 完整的啮齿动物和人体组织的广泛分子解剖学。在第一阶段的提案中，半透明生物系统公司 将建立Clearbot™的原型，并展示用于全脑分析的自动脑组织清除。 具体目标：目标1：组装和评估Clearbot原型：原型的基本功能 将对工程顾问开发的功能、稳健性、能耗和 整体可靠性。目的2：评价Clearbot用于自动清除和免疫染色的实用性。 完整的大脑：将使用Aim中组装的原型评价组织清除的质量和可重复性 1.将清除多个小鼠大脑，并使用Clearbot进行免疫染色，并以手动进行基准测试 并行处理。这个第一阶段项目将证明开发一个自动化的 与手动清除相当或更好的设备。在第一阶段可行性测试成功之后， 未来的发展将把原型推进到一个灵活的、商业级的系统， 用户界面，帮助研究人员将这种改变游戏规则的技术应用于他们的研究问题。

SUMMARY Tissue clearing is a powerful technique that renders large biological samples optically transparent and allows 3D visualization of wide-brain molecular anatomy. To help disseminate this technology, Translucence Biosystems developed a Mesoscale Imaging System™ and, using previous SBIR NIH support, an analytical software package for advanced analysis of brain samples, 3TKTM. While complete solutions exist for imaging and analysis of cleared samples, no complete solution exists for the highly labor intensive and error prone tissue clearing process. Here Translucence Biosystems introduce Clearbot™, an automated system for sample clearing and immunostaining that will complement previous Translucence devices and complete a pipeline for reliable brain- wide molecular anatomy of intact rodent and human tissues. In this Phase I proposal, Translucence Biosystems will build a prototype of Clearbot™ and demonstrate automated brain tissue clearing for whole-brain analysis. Specific aims: Aim 1: Assemble and evaluate a Clearbot prototype: the basic functionality of the prototype developed by an engineering consultant will be evaluated for functionality, robustness, energy consumption, and overall reliability. Aim 2: Evaluate the utility of Clearbot for automated clearing and immunostaining of intact brains: quality and repeatability of tissue clearing will be evaluated using the prototype assembled in Aim 1. Multiple mouse brains will be cleared and immunostained with Clearbot and benchmarked to manual processing done in parallel. This Phase I project will demonstrate the feasibility of developing an automated device that is comparable to or better than manual clearing. Following success of this Phase I feasibility testing, future developments will advance the prototype to a flexible, commercial-grade system with a robust and simple user-interface, helping researchers apply this game-changing technology to their research questions.