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 组织清除是一种强大的技术，它使大型生物样品在光学上透明并允许3D 大脑分子解剖结构的可视化。为了帮助传播这项技术，半透明生物系统 开发了中尺度成像系统™，并使用先前的SBIR NIH支持，一种分析软件 用于大脑样品的高级分析的包装，3TKTM。虽然存在用于成像和分析的完整解决方案 在清除的样品中，不存在针对高度强度和误差组织清除的完整解决方案 过程。在这里，《半透明生物系统》介绍了Clearbot™，这是一种用于样品清除和的自动化系统 免疫染色，将完成以前的半透明设备，并完成一条管道，以实现可靠的大脑 完整的啮齿动物和人体组织的宽分子解剖结构。在此阶段I建议中，半透明生物系统 将构建Clearbot™的原型，并展示自动化的脑组织清除，以进行全脑分析。 具体目的：目标1：组装和评估Clearbot原型：原型的基本功能 由工程顾问开发的功能，鲁棒性，能耗和 总体可靠性。目标2：评估Clearbot自动清除和免疫染色的实用性 完整的大脑：将使用AIM组装的原型评估组织清除的质量和可重复性 1。多个小鼠的大脑将被清除并通过Clearbot进行免疫染色，并根据手册进行基准测试 并行进行处理。该阶段I项目将证明开发自动化的可行性 与手动清除相当或更好的设备。在此阶段I的成功之后，我可行性测试， 未来的发展将以强大而简单的方式将原型推向灵活的商业级系统 用户界面，帮助研究人员将这种改变游戏的技术应用于他们的研究问题。