Ultrahigh Resolution Optical Barcode

超高分辨率光学条形码

• 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
• 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

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）：随着对进行更高通量和更低成本反应的期望的增加，多路复用技术在生物学和生物化学领域中已经稳步普及。光学多路复用包括将已知的生物探针连接到具有已知光学代码的珠上，其允许通过阅读其独特的光学代码来光学识别合并样品中的每个探针。在随后的步骤中，染料标记的靶标的发射与探针身份相关联，以确定所讨论的反应的程度。目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，但目前只有<100种光学代码可商购获得（例如来自Luminex）。对于第一阶段的努力，我们假设，如果用窄发射体如稀土元素代替有机染料的宽发射（在FWHM处高达30-60 nm），则有可能获得更高分辨率的光码，所述稀土元素通常显示2-10 nm范围内的发射峰宽度。在第二阶段，我们证明了在理想条件下，通过使用已知量的稀土基Parallume材料，可以分辨多达10亿个光码，这些材料可以发射多达6种颜色的光多路复用。该系统相对于当前基于流式细胞仪的系统的优点包括：来自每个发射器的光谱离散发射、通过使用可变发射器浓度可获得的非常高水平的多路复用、高量子效率、优异的光稳定性、可变颗粒尺寸以及低成本、自动化并行合成。非常重要的是，我们还开发了一种完全便携式的电池供电的原型微珠读取器（多路检测读取器系统，或“MARS”），其具有基于非常便宜的商用DSLR相机的成像系统的超亮LED。为了响应NHGRI赞助的复杂仪器竞争更新应用程序的特定计划公告PA-08-115，并行合成建议基于由Parallume珠，MARS硬件和软件组成的Parallume平台构建一个完整的后期原型。具体地，我们提出（1）开发用于Parallume珠的自动化、并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MARS原型并将它们递送给合作者进行beta测试，从而产生反馈以创建原型的最终版本，（3）创建用于MARS中的低成本珠定位载玻片（BLS）以实现高密度珠图像，（4）完成软件包以控制MARS并分析从MARS输出的数据，也将与MARS沿着进行β测试，以及（5）通过将我们的恰加斯病诊断测定与使用Luminex的已知诊断测定进行比较来测试完全功能性的系统。在三年的开发期结束时，并行合成将有一个功能齐全的G3 MARS与用户友好的软件沿着随着BLS和编码的Parallume珠作为消耗品。该平台为多重检测开发和复杂生物成像和可视化领域提供了全新的技术，将提供给潜在的战略合作伙伴、被许可方或直接销售给最终用户。这种低成本系统非常适合低资源设置应用，如发展中国家。公共卫生关系：如果可以同时进行许多反应，那么发达国家和低资源环境中使用的测定将大大受益于降低的成本和增加的通量。为了一次处理许多混合样品，需要有区分单个样品的方法。该提议描述了一种方法，通过该方法，每个珠在用激光激发时发射唯一的光学特征。每个微珠的光学特征允许测定微珠的含量和测定期间的反应程度。通过这种方法可以解析成千上万的光码。通过便携式检测器/微珠读取器阅读光学特征，可以廉价地并行执行和分析数千次测定。