Multi-color laser particles for high-throughput pooled analysis

摘要 大量样品的高通量分析可以大大受益于采样 条形码，因为它可以显着降低试剂成本和批次错误， 采集速度光学条形码的读数与荧光分析兼容， 与DNA条形码相比，它具有令人信服的优势，因为它允许快速的多参数分析， 每个样本，并且不依赖于二进制的“命中”或“未命中”标准。然而，目前的荧光- 基于条形码的方法不能满足当前和未来的大规模复用的需要。的 该项目的目标是开发新的，高度可扩展的，光复用技术的基础上， 多色激光粒子。几个盘状微粒的组合，每个微粒发射一个 窄带激光光谱，可以标记在细胞或微珠的表面上， 对象的唯一标识符。第一个具体目标是开发50套彩色 半导体激光器粒子通过微流体粒子在1150至1600 nm的光谱范围内 分类第二个具体目标是开发这种新工具在三个主要领域的应用， 通量分析平台：首先，384个混合细胞样品的多参数流式细胞术; 第二，3840重珠免疫测定;第三，全光学多参数筛选， 组合一珠一化合物文库。这些高通量的突破 技术有望促进许多生物发现应用，包括药物 发展

Abstract High-throughput analysis of a large number of samples can greatly benefit from sampling barcoding as it can significantly reduce reagent costs and batch error as well as increasing the acquisition speed. Optical barcoding with its readout compatible with fluorescence analysis offers compelling advantages over DNA barcoding because it allows rapid multiparameter analysis of each sample and does not rely on a binary ‘hit’ or ‘miss’ criterion. However, current fluorescence- based barcoding does not meet the current and future needs of massive-scale multiplexing. The goal of this project is to develop novel, highly scalable, optical multiplexing technology based on multi-color laser particles. A combination of several discoidal microparticles, each emitting a narrowband laser spectrum, can be tagged on the surface of cells or microbeads and serve as the unique identifiers of the objects. The first specific aim is to develop 50 color sets of semiconductor laser particles across a spectral range of 1150 to 1600 nm via microfluidic particle sorting. The second specific aim develops applications of this new tool in three major high- throughput analysis platforms: first, multiparameter flow cytometry of 384-pooled cell samples; second, 3840-plex bead immunoassays; and third, all-optical multiparameter screening of combinatorial one-bead-one-compound libraries. The breakthroughs in these high-throughput technologies are expected to facilitate numerous biological discovery applications including drug development.