NEXT GENERATION OPTICALLY ACTIVATED LARGE PARTICLE SORTING

下一代光激活大颗粒分选

• 批准号: 8361769
• 负责人: STEVEN W GRAVES
• 金额: $ 8.95万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361769
• 关键词: Caliber; Cell Separation; Charge; Chromosomes; Collaborations; Collection; Coupled; Development; Event; Flow Cytometry; Funding; Grant; Lasers; Libraries; Mechanics; Molecular; National Center for Research Resources; Optics; Particle Size; Peptide aptamers; Positioning Attribute; Principal Investigator; Research; Research Infrastructure; Resources; Solutions; Sorting - Cell Movement; Source; Speed; Stream; System; Techniques; Technology; Testing; Uncertainty; United States National Institutes of Health; Whole Organism; base; combinatorial; cost; data acquisition; next generation; particle; prevent; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Several avenues of research have a critical need for high speed sorting of large particles, including the selection of whole organisms, multicellular particles and the selection of molecules from combinatorial libraries synthesized on large particles. High speed sorting has to date primarily relied on charge based droplet sorting coupled with multiparameter optical analysis of particles in a flow cytometer, which is an indispensable technique for numerous biomedical applications including rare cell isolation, chromosome sorting and cellular display molecular selection among many others. However, droplet based flow sorters have significant limitations when large particles are considered. First, increasing particle size requires largerorifices to prevent clogging and effects on droplet break off points. Sorting orifices suffer increasing turbulence as their diameter increases, which requires the use of lower linear velocities and restricts sorting rates (<1000/s). This, effectively limits particle size (<100 ¿m) and has led to alternative large particle sorting approaches, including mechanical stream diversion and micro-channel fluidic switching. Our effort focuseson solutions to large particle sorting that will result in the development of sorters that will sort particles up to 1 mm in diameter at rates comparable to current conventional droplet based sorters (>104/s). To accomplish this, we will target each technical limitation of current sorting technology with unique solutions for large particle systems. First, we will develop high speed synchronous particle delivery systems to overcome the statistical uncertainty of particle delivery and maximize sorting rates by providing known particle positions for sorting events. Second, we will leverage our recent low cost flow cytometry developments in lasers and data acquisition systems to create inexpensive low linear velocity parallel flow cytometry analyzers to maximize particle throughput. Third, we will create droplet on demand sorters that are not limited by particle size. While these technical developments will be most applicable to large particle sorting, they will also have ancillary benefits to conventional sorting, as they will dramatically reduce system cost and create valuable parallel analysis technology for high speed sorting. Finally, we will construct high-speedlarge particle sorters for internal testing and key external collaborations to sort large particles at high rates for selection of aptamers and peptides as well as rare tumor microspheroid collection.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 多种研究途径迫切需要对大颗粒进行高速分选， 包括整个生物体、多细胞颗粒和分子的选择 来自在大颗粒上合成的组合文库。迄今为止，高速分拣已主要 依赖于基于电荷的液滴分​​选以及对颗粒的多参数光学分析 流式细胞仪是众多生物医学应用中不可或缺的技术，包括 稀有细胞分离、染色体分选和细胞显示分子选择等。 然而，当大颗粒被排出时，基于液滴的流式分选机具有显着的局限性。 经过考虑的。首先，增加颗粒尺寸需要更大的孔口以防止堵塞和影响 液滴断裂点。随着分选孔直径的增加，其紊流也会增加， 这需要使用较低的线速度并限制分选速率（<1000/s）。这， 有效限制颗粒尺寸（<100 µm）并导致替代性大颗粒分选方法， 包括机械流转向和微通道流体切换。我们的努力集中于 大颗粒分选的解决方案将导致分选颗粒的分选机的开发 直径高达 1 毫米，速度与当前基于传统液滴的分选器 (>104/s) 相当。 为了实现这一目标，我们将针对当前分拣技术的每个技术限制 大颗粒系统的独特解决方案。首先，我们将开发高速同步粒子 输送系统克服颗粒输送的统计不确定性并最大化分选 通过为排序事件提供已知的粒子位置来确定速率。其次，我们将利用最近的 低成本流式细胞仪在激光器和数据采集系统方面的发展，创造了廉价的 低线速度平行流式细胞术分析仪可最大限度地提高颗粒通量。第三，我们将 创建不受颗粒尺寸限制的按需液滴分选机。虽然这些技术 发展将最适用于大颗粒分选，它们还将具有附带好处 与传统排序相比，它们将大大降低系统成本并创造有价值的并行 高速分选分析技术。最后，我们将构建高速大粒子 用于内部测试和关键外部合作的分选机，以高速率分选大颗粒 适体和肽的选择以及罕见的肿瘤微球体的收集。