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

Clearbot：全自动、高通量组织透明化和免疫染色系统

• 批准号: 10382515
• 负责人: Damian G. Wheeler
• 金额: $ 45.97万
• 依托单位: TRANSLUCENCE BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-16 至 2025-02-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10382515
• 关键词: 3-Dimensional; Accounting; Address; Adoption; Agitation; Anatomy; Automation; BRAIN initiative; Behavioral Research; Benchmarking; Biological; Brain; Brain imaging; Cell Count; Chemicals; Complement; Computer software; Computer-Aided Design; Consumption; Corrosives; Data Analyses; Development; Devices; Ecosystem; Encapsulated; Engineering; Exposure to; Funding Opportunities; Future; Growth; Histologic; Image; Imaging technology; Legal patent; Light; Liquid substance; Manuals; Methodology; Methods; Microscope; Molecular; Mus; Neurosciences; Noise; Optics; Organ; Parvalbumins; Phase; Predisposition; Procedures; Process; Protocols documentation; Reagent; Reporting; Reproducibility; Reproducibility of Results; Research; Research Personnel; Risk; Rodent; Sampling; Signal Transduction; Small Business Innovation Research Grant; Solvents; Stains; System; Techniques; Technology; Temperature; Testing; Time; Tissues; United States National Institutes of Health; Validation; Visualization; Work; base; brain research; brain tissue; commercialization; data management; design; drug development; feasibility testing; flexibility; human error; human tissue; imaging system; improved; instrumentation; model design; prototype; research and development; software systems; success; three-dimensional visualization; tissue processing; tool

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.

概括 组织透明化是一种强大的技术，可以使大型生物样本光学透明并允许 3D 为了帮助传播这项技术，Translucence Biosystems 开发了宽脑分子解剖学的可视化。 开发了 Mesoscale Imaging System™，并利用之前的 SBIR NIH 支持开发了分析软件 用于大脑样本高级分析的软件包 3TKTM，同时存在用于成像和分析的完整解决方案。 对于已清除的样品，对于高度劳动密集型且容易出错的组织清除，不存在完整的解决方案 Translucence Biosystems 在此推出 Clearbot™，这是一种用于样品清理和处理的自动化系统。 免疫染色将补充之前的 Translucence 设备，并完成可靠的脑-免疫染色的管道 在此第一阶段的提案中，Translucence Biosystems 对完整的啮齿动物和人体组织进行了广泛的分子解剖。 将构建 Clearbot™ 原型并演示用于全脑分析的自动脑组织清除。 具体目标： 目标 1：组装并评估 Clearbot 原型：原型的基本功能 由工程顾问开发的产品将对其功能、稳健性、能耗和性能进行评估 目标 2：评估 Clearbot 在自动清除和免疫染色方面的实用性。 完整的大脑：将使用 Aim 中组装的原型来评估组织透明化的质量和可重复性 1. 使用 Clearbot 清除多只小鼠大脑并进行免疫染色，并以手动为基准 该第一阶段项目将展示开发自动化系统的可行性。 在第一阶段可行性测试成功后，该设备可与手动清算相媲美或更好， 未来的发展将把原型推进到一个灵活的商业级系统，具有强大而简单的功能 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。