Nanoparticle Tracking Analyzer (NTA) for the Center for Live Cell Genomics

主要研究者/项目负责人：Haussler，大卫 项目摘要 我们的CEGS中心活细胞基因组学的重点是建立新的方法和能力， 大规模、长期、廉价、模块化、可定制、共享、物联网控制、可复制 活细胞培养和基于组织的实验基因组学疾病模型。组织模型包括传统的细胞 培养物以及从手术或活组织检查获得的类器官和原代组织外植体。一个特别的重点是 支持组织生长和维持的类器官工厂与外部和芯片上的集成 电光流控分析模块成为生态系统的一部分， 软件该系统将使用商品传感器，摄像机和计算机连接在灵活的平台上， 设计使用简单，广泛可用的技术，以刺激快速创新， 组织培养的实验平台。这项新技术将使我们能够解决重大的科学问题 在神经发育和儿童癌症中的作用。这些问题包括什么基因有助于人类 大脑发育，或者在个别儿科癌症病例中哪些特定的分子途径被破坏。 在我们项目的第一年，很明显，为了推进我们在两个方面的研究议程， 在科学和技术方面，我们需要一种已建立的多模式纳米颗粒分析方法。 具体而言，我们面临两个挑战。我们需要能够评估纳米级的细胞外囊泡 (EVs)，特别是外泌体，它们在我们的类器官工厂中产生，作为衡量类器官细胞 类型、新陈代谢和健康。由于这些测量需要在长时间内频繁进行， 因此，需要一种快速且相对易于使用的技术。其次，为了开发光流体器件， 可以对单个EV和外泌体及其分子含量进行测量，我们需要 能够建立地面实况参考，这些设备的性能可以与之进行比较。到 为此，我们正在申请补充资金，以获得纳米粒子跟踪分析仪（NTA） 这些工具将在我们的整个项目中提供这些功能。NTA是一种独特的工具， 提供生物和无机纳米粒子群体的多模态分析。相关能力 包括缓冲液校准、EV浓度和尺寸分布测量、表面电荷 测量、标签效率评估等等。因此，NTA是唯一适合我们的 这将大大加快我们的研究进展，并扩大其成果。 1

Principal Investigator/Program Director: Haussler, David Project Summary The focus of our CEGS Center for Live Cell Genomics is to build new methodology and capacity for large-scale, long-term, inexpensive, modular, customizable, shared, Internet-of-Things- controlled, reproducible live cell culture and tissue-based experimental genomics disease models. Tissue models include traditional cell culture as well as organoid and primary tissue explants obtained from surgery or biopsy. A particular focus is the integration of organoid factories that support tissue growth and maintenance with external and on-chip electro- optofluidic analytical modules to become part of an ecosystem that is modeled after open- source software. This system will use commodity sensors, cameras, and computers linked in platforms that are flexibly designed using simple, widely available techniques potentially in order to stimulate rapid innovation in experimental platforms for tissue culture. This novel technology will allow us to address major scientific issues in neurodevelopment and pediatric cancer. These include questions about what genes contribute to human brain development, or what specific molecular pathways are disrupted in individual pediatric cancer cases. Over the first year of our project, it has become clear that in order to advance our research agenda on both the scientific and technological sides we require an established, multi-modal nanoparticle analysis method. Specifically, we are facing two challenges. We need to be able to assess the nanoscale extracellular vesicles (EVs), specifically exosomes, that are being produced in our organoid factories as a measure of organoid cell types, metabolism, and health. As these measurements need to be carried out frequently over long periods of time, a fast and relatively easy-to-use technique is required. Secondly, in order to develop optofluidic devices that can carry out measurements on individual EVs and exosomes and their molecular content, we need to be able to establish ground truth references against which the performance of these devices can be compared. To this end, we are requesting supplemental funding to acquire a Nanoparticle Tracking Analyzer (NTA) instrument that will provide these capabilities throughout our project. An NTA is a unique instrument that provides multi-modal analysis on populations of biological and inorganic nanoparticles. Pertinent capabilities include buffer calibration, measurement of EV concentration and size distribution, surface charge measurements, assessment of labeling efficiencies, and more. As such, an NTA is uniquely suited for our requirements and will dramatically accelerate the progress of our research as well as expand its outcomes. 1