Rapid Evaluation of Neuronal Activity in the Intact Whole Brain at Single Cell Resolution

以单细胞分辨率快速评估完整全脑的神经元活动

• 批准号: 10630313
• 负责人: Damian G. Wheeler
• 金额: $ 40.05万
• 依托单位: TRANSLUCENCE BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10630313
• 关键词: Anatomy; BRAIN initiative; Basic Science; Behavior; Biological Assay; Brain; Brain imaging; Calcium; Cells; Code; Communities; Computer software; Coupled; Data; Detection; Development; Diagnostic Reagent Kits; Evaluation; FOS gene; Fluorescence; Gene Proteins; Genes; Image; Imaging technology; Immediate-Early Genes; Individual; Knowledge; Light; Link; Maps; Marketing; Measures; Mus; NPAS4 gene; Neurons; Neurosciences; Optics; Performance; Phase; Population; Procedures; Process; Production; Reagent; Regulation; Research Personnel; Resolution; Scientist; Services; Small Business Innovation Research Grant; Software Tools; Speed; Stains; Stimulus; Strategic vision; Technology; Testing; Therapeutic; Tissues; Work; activity marker; area striata; computerized tools; design; diagnostic platform; drug development; fluorescence imaging; head-to-head comparison; improved; interest; machine learning framework; microscopic imaging; neural; pre-clinical; pre-clinical research; recruit; response; statistics; tool; two-photon; user friendly software; visual stimulus

Project Summary Here we seek to develop a “Neuronal Activity Quantitative Diagnostic” platform and reagent kits to disseminate accessible products for measuring recent neural activity across the entire mouse brain to the neuroscience community. Previously we have shown that the activity-dependent immediate-early gene protein products Npas4 and cFos can be clearly detected, imaged, and quantified using developmental versions of our platform tools. Immediate early genes are genes that are activated transiently and rapidly in response to a wide variety of cellular stimuli. The three overlapping aims to be investigated in this Phase II project are: 1) Demonstration of the fidelity of our assay for our customers. To demonstrate the robustness and accuracy of our neuronal activity assay with another well-known assay for activity, we will perform two photon calcium fluorescence imaging in the primary visual cortex of mice presented with repeated visual stimuli. We will clear and stain the brains for cFos and Npas4 and demonstrate the fidelity of these markers for detecting recently active neurons. 2) Develop and beta test robust versions of our software tools useable by researchers with no specialized coding or statistics knowledge. We will develop Stitchy and BrainSnap into robust, user-friendly software products that remove the current analysis bottleneck for neuroscientists interested in whole-brain imaging. Stitchy will be an improved browser-based version of our internal software that simplifies and speeds the process of stitching together light sheet microscope images. BrainSnap software will integrate our high-performance 3TK- Quant framework for machine learning-enabled quantification and our statistical framework for whole-brain anatomics, 3TK-Stats, in a browser-based interface. We have recruited end users who will test this software in a head-to-head comparison with our in-house testing. 3) Develop and test a brain clearing and staining kit for neuronal activity. To help researchers produce high quality cleared brains stained with neuronal activity markers, we will generate Neuronal Activity clearing and staining reagents kits for cFos and Npas4. These will be tested by collaborators in head-to-head comparisons with our in-house procedures on brains prepared in parallel. At the end of Phase II, Translucence will have a market-ready Neuronal Activity Quantitative Diagnostic platform including reagent kits, user-friendly software, and service options.

项目摘要 在这里，我们试图开发一个“神经元活动定量诊断”平台和试剂盒 传播可访问的产品，用于测量整个小鼠大脑的最新神经活动 神经科学社区。以前我们已经表明活性依赖于立即自耳的基因蛋白 可以使用我们的发育版本清楚地检测，成像和量化产品NPAS4和CFO 平台工具。早期基因的立即是基因，这些基因响应于宽阔的基因 各种细胞刺激。在此II阶段项目中要调查的三个重叠目标是： 1）证明我们对客户评估的保真度。展示鲁棒性和准确性 我们的神经元活性评估以及另一种众所周知的活动评估，我们将执行两个光子钙 用反复的视觉刺激呈现的小鼠的一级视觉皮层中的荧光成像。我们会清除 并染色CFO和NPAS4的大脑，并证明了这些标记的忠诚度最近检测 活性神经元。 2）开发和beta测试我们的软件工具的强大版本，由没有专业的研究人员可用 编码或统计知识。我们将开发缝线和大脑，为强大的，用户友好的软件 消除当前分析瓶颈的产品，这些神经科学家对全脑成像感兴趣。 Stitchy将是我们内部软件的基于浏览器的改进版本，可简化和加快流程 将轻板显微镜图像缝合在一起。 BrainsNap软件将整合我们的高性能3TK- 用于机器学习的定量量化量子框架和我们全脑的统计框架 基于浏览器的界面中的解剖学，3TK-stats。我们招募了最终用户，他们将在 与我们的内部测试的正面比较。 3）开发和测试用于神经元活性的大脑清除和染色试剂盒。帮助研究人员产生高 用神经元活动标记染色的质量清除大脑，我们将产生神经元活动清除和 CFO和NPAS4的染色试剂盒。这些将由合作者在正面比较中测试 我们的内部手术并行准备大脑。 在第二阶段结束时，半透明将具有市场就绪的神经元活动定量诊断 包括试剂套件，用户友好的软件和服务选项的平台。