Ultrahigh Resolution Optical Barcode

超高分辨率光学条形码

基本信息

批准号：
7915727
负责人：
ROBERT C HAUSHALTER
金额：
$ 74.81万
依托单位：
PARALLEL SYNTHESIS TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-26 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7915727
关键词：
Algorithms Automation Biochemistry Biological Biological Assay Biology Blood California Chagas Disease Characteristics Code Color Complex Computer software Custom Data Detection Developed Countries Developing Countries Development Devices Diagnosis Diagnostic Dyes Evaluation Feedback Film Generations Goals Housing Image Individual Infection Injection of therapeutic agent Label Laboratories Lasers Liquid substance Manuals Measures Methodology Methods Molds NIH Program Announcements National Human Genome Research Institute Optics Output Parasites Particle Size Performance Phase Process Proteins Quantum Dots Rare Earth Metals Reaction Reader Reading Recombinants Reporter Reproducibility Research Research Design Resolution Resources Sales Sampling San Francisco Series Signal Transduction Silicon Site Slide Staging System Systems Analysis Technology Test Result Testing Trypanosoma cruzi Universities Width assay development base computerized data processing cost density design detector functional group image processing image visualization instrument instrumentation new technology point of care point-of-care diagnostics prevent prototype public health relevance quantum ratiometric research study response software development software systems user friendly software

项目摘要

DESCRIPTION (provided by applicant): Multiplexing technologies have steadily gained in popularity in the fields of biology and biochemistry as the desire to perform higher-throughput and lower-cost reactions has increased. Optical multiplexing consists of attaching a known biological probe to a bead with a known optical code which allows the optical identification of each probe in the pooled sample by reading its unique optical code. In a subsequent step, the emission of a dye-labeled target is associated with the probe identity to determine the extent of the reaction in question. The two classes of materials currently used for optical multiplexing are fluorescent organic dyes and Quantum Dots but only <100 optical codes are currently available commercially (e.g. from Luminex). For the Phase I effort we hypothesized that it would be possible to obtain far more resolvable optical codes if the broad emission from organic dyes (up to 30-60 nm at FWHM) were replaced with narrow emitters such as the rare earth elements which often display emission peak widths in the 2-10 nm range. In Phase II we demonstrated that it was possible to resolve up to a billion optical codes under ideal conditions by using known amounts of rare-earth-based Parallume materials which emit up to six colors for optical multiplexing. The advantages of this system over the current flow cytometer-based systems include: spectrally discrete emission from each of the emitters, a very high level of multiplexing available through the use of variable emitter concentrations, high quantum efficiency, excellent photostability, variable particle size, and a low cost, automated parallel synthesis. Very importantly, we have also developed a completely portable, battery-powered prototype bead reader (Multiplexed Assay Reader System, or "MARS") with on superbright LEDs with an imaging system based on a very inexpensive commercial DSLR camera. In response to the specific Program Announcement, PA-08-115, for Competing Renewal applications for Complex Instrumentation under the auspices of the NHGRI, Parallel Synthesis proposes to build a complete late-stage prototype based on the Parallume platform consisting of the Parallume beads, the MARS hardware and software. Specifically, we propose to (1) develop an automated, parallel synthesis methodology for the Parallume beads to fulfill encoded set requirements, (2) create two automated, second-generation MARS prototypes and deliver them to collaborators for beta- testing resulting in feedback to create a final version of the prototype, (3) create a low-cost bead localization slide (BLS) for use in the MARS to enable high-density bead images, (4) complete a software package to control the MARS and analyze the data output from the MARS, also to be beta-tested along with the MARS, and (5) test the completely functional system by comparing our diagnostic assay for Chagas disease against a known diagnostic assay using Luminex. At the end of the three-year development period, Parallel Synthesis will have a fully functional G3 MARS with user- friendly software along with the BLS and encoded Parallume beads as consumables. This platform, offering a completely new technology for the fields of multiplex assay development and complex biological imaging and visualization, will be available for potential strategic partners, licensees, or for direct sale to end-users. This low cost system is ideally suited for low resource setting applications such as developing countries. PUBLIC HEALTH RELEVANCE: Assays used in both the developed countries and low resource settings would greatly benefit from reduced costs and increased throughput if it were possible to perform many reactions simultaneously. In order to process many pooled samples at once it is necessary to have a means of distinguishing the individual samples. This proposal describes a method by which each bead emits a unique optical signature upon excitation with a laser. The optical signature of each bead allows the determination of the bead's content and the extent of reaction during the assay. It is possible to resolve many thousands of optical codes by this method. Reading of the optical signatures by a portable detector/bead reader allows thousands of assays to be inexpensively performed and analyzed in parallel.

描述（由申请人提供）：随着对进行高通量和低成本反应的渴望，多重技术在生物学和生物化学领域稳步越来越受欢迎。光学多路复用包括将已知的生物学探针连接到具有已知光学代码的珠子上，该探针通过阅读其唯一的光学代码，可以在汇总样品中对每个探针进行光学识别。在随后的步骤中，染料标记靶标的发射与探针身份相关，以确定所讨论的反应程度。当前用于光学多路复用的两类材料是荧光有机染料和量子点，但目前在商业上仅可用<100个光学代码（例如，来自Luminex）。在第一阶段的努力中，我们假设如果有机染料的广泛发射（在FWHM最高30-60 nm）被狭窄的发射器（例如稀土元素）取代，那么可以获得更多可分解的光学代码。 2-10 nm范围内的发射峰宽度。在第二阶段中，我们证明，在理想条件下，可以使用已知数量的基于稀土的派别派别材料来解决多达十亿个光学代码，这些拼写材料最多可发出六种颜色进行光学多重。该系统比基于当前流式细胞仪的系统的优点包括：每个发射器的光谱离散发射，通过使用可变的发射极浓度，高量子效率，出色的光稳定性，可变粒径，可变粒径，可变的多重级别，可用以及低成本的自动平行合成。非常重要的是，我们还开发了一个完全可移植的，电池供电的原型珠读取器（多路复用测定读取器系统或“火星”），并在Superbright LED上使用基于非常便宜的商业DSLR相机的成像系统进行了成像系统。为了响应特定的计划公告，PA-08-115，用于在NHGRI的主持下为复杂仪器进行竞争的续订应用程序，并行合成建议建立一个基于由派系珠的派别平台，由派系珠组成的完整阶段原型火星硬件和软件。具体而言，我们建议（1）为阶段珠开发一种自动化的平行合成方法，以满足编码的集合要求，（2）创建两个自动化的，第二代火星原型，并将其交付给合作者，以进行β-测试，从而导致反馈以进行反馈至创建原型的最终版本，（3）创建一个低成本的珠子定位幻灯片（BLS），以供火星使用以启用高密度珠图像，（4）完成一个软件包来控制火星并分析数据火星的输出，也将与火星一起进行β测试，（5）通过比较我们对chagas疾病的诊断测定法与使用Luminex的已知诊断测定法测试完全功能性的系统。在三年开发期结束时，并行合成将具有功能齐全的G3火星，并具有用户友好的软件以及BLS和编码的众多珠子作为消耗品。该平台将为多重测定开发和复杂的生物成像和可视化领域提供全新的技术，可用于潜在的战略合作伙伴，被许可人或直接向最终用户销售。这种低成本系统非常适合低资源设置应用，例如发展中国家。公共卫生相关性：在发达国家使用的测定和资源较低的测定能力将从降低的成本和增加的吞吐量中受益匪浅，如果可以同时执行许多反应。为了立即处理许多合并样品，有必要有一种区分各个样本的方法。该提案描述了一种方法，每个珠子在用激光激发时会发出独特的光学特征。每个珠的光学特征允许测定珠的含量和测定过程中的反应程度。通过这种方法可以解决数千种光学代码。通过便携式检测器/珠读取器读取光学特征，可以并行进行数千种测定法。