Ultrahigh Resolution Optical Barcode

描述(申请人提供)：第一阶段工作的中心假设是，如果能够合成一种能够发出数百万独特的、可分辨的光学特征的材料，并且诸如DNA的生物分子能够牢固地附着在它们上面，那么这种深度多路复用的珠集可以形成“悬浮阵列”的基础，其中每个编码粒子的光学指示符唯一地识别给定的珠子。一旦每一颗珠子都可以通过其光学代码进行识别，那么高度平行的反应就成为可能，比如一次杂交数百万个样本。六元发射体Y1-x[DyaErbEucHodSmeTmf]xVO4在450-700 nm范围内有六个窄的发射峰，当YVO4主体被激发到325 nm(He-Cd激光)时，根据六个发射体的相对积分强度显示出数百万种独特的光学特征。对大量样品的统计分析表明，大约2x10(9)个独特的光学代码可以被解析，因为六个发射器中的一个作为内部标准而被“牺牲”，从而能够执行非常精确和精确的比率测量。实验表明，Cy3标记的DNA可以牢固地附着在编码可控孔玻璃(CPG)或纯YVO4上，来自有机杂交染料报告的信号和无机光学代码可以在单个粒子内相互独立地读取。第二阶段的工作将以一种基于热蜡转印的新的超高通量珠子制造方法生产光学编码材料，预计每小时可生产500-5000万个编码颗粒。在适当的衍生化和将DNA或蛋白质附着到珠子之后，将在烧瓶中集体进行各种杂交和蛋白质结合实验。在将编码的颗粒放置在防止颗粒重叠的特殊衬底上之后，使用Parallel的高光谱成像系统随后进行解码，将从采集的图像中的每个像素产生可见的发射光谱，从而提供了在空间上测量多色发射的非常高吞吐量的手段。该技术的精确度、吞吐量和成本估算表明，与目前基于高密度芯片的基因分型和基因表达分析相比，它具有很强的竞争力。 这项正在开发中的技术将允许一种独特的光学代码附着在生物分子上，从而使它们能够被数百万次分析。对大量小样本的并行准确光学分析将降低成本，增加可用于多种类型医学样本的样本数量。

DESCRIPTION (provided by applicant): The central hypothesis of the Phase I effort was that if a material could be synthesized that could emit millions of unique, resolvable optical signatures, and biomolecules such as DNA could be firmly attached to them, that this deeply multiplexed bead set could form the basis of a "suspension array" where the optical designator of each encoded particle uniquely identifies a given bead. Once each bead is identifiable via its optical code, then highly parallel reactions, like hybridization of millions of samples at once, become possible. The hexanary emitter Y1-x[DyaErbEucHodSmeTmf]xVO4, which has six narrow emission peaks in the 450-700nm range, displays millions of unique optical signatures based on the relative integrated intensities of the six emitters when the YVO4 host is excited at 325nm (He-Cd laser). The statistical analysis of a large number of samples showed that approximately 2 x 10(9) unique optical codes could be resolved, because one of the six emitters is "sacrificed" as an internal standard allowing very accurate and precise ratiometric measurements to be performed. Experiments have shown that cy3-labeled DNA could be firmly attached to either encoded Controlled Pore Glass (CPG) or pure YVO4 and the signals from the organic hybridization dye reporters and the inorganic optical code can be read independently from one another within a single particle. The Phase II effort will produce the optically encoded materials in a new super high throughput bead fabrication method based on thermal wax transfer printing where it is estimated that 5-50 million encoded particles per hour can be produced. After appropriate derivitization and attachment of DNA or proteins to the beads, a variety of hybridization and protein binding experiments will be performed en masse in a flask. Subsequent decoding using Parallel's hyperspectral imaging system, after placement of the encoded particles onto a special substrate which prevents overlap of the particles, will produce a visible emission spectrum from each pixel in the acquired image providing a very high throughput means of spatially measuring the multicolor emission. The accuracy, throughput and cost estimated for this technology indicate that it could be very competitive with the current high density chip-based assays for genotyping and gene expression. The technology under development will allow a unique optical code to be attached to biomolecules thereby allowing them to be analyzed millions at time. The accurate optical analysis of very large numbers of small samples in parallel will reduce the cost, and increase the number of samples that can be examined, for many types of medical samples.