Ultrahigh Resolution Optical Barcode

描述（由申请人提供）：第一阶段工作的中心假设是，如果可以合成一种材料，该材料可以发出数百万个独特的、可解析的光学特征，并且生物分子（如DNA）可以牢固地附着在它们上面，那么这种深度复用的头集可以形成“悬浮阵列”的基础，其中每个编码粒子的光学指示符唯一地标识给定的头。一旦每个珠子都可以通过光学编码识别出来，那么高度平行的反应，比如同时进行数百万个样本的杂交，就成为可能。六元发射体Y1-x[DyaErbEucHodSmeTmf]xVO4在450 ~ 700nm范围内具有6个窄发射峰，当YVO4主体在325nm （He-Cd激光）激发时，基于6个发射体的相对集成强度，显示出数百万个独特的光学特征。大量样品的统计分析表明，大约2 × 10(9)个独特的光学代码可以被解决，因为六个发射器中的一个被“牺牲”作为内部标准，允许非常准确和精确的比率测量进行。实验表明，cy3标记的DNA可以牢固地附着在编码的受控孔玻璃（CPG）或纯YVO4上，并且来自有机杂交染料报告器和无机光学代码的信号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移印刷的新型超高通量珠制造方法生产光学编码材料，估计每小时可以生产500 - 5000万个编码颗粒。在适当的衍生化和DNA或蛋白质附着到珠上后，将在烧瓶中大量进行各种杂交和蛋白质结合实验。随后使用Parallel的高光谱成像系统进行解码，在将编码的粒子放置在防止粒子重叠的特殊衬底上之后，将从获得的图像中的每个像素产生可见的发射光谱，提供空间测量多色发射的非常高的吞吐量手段。该技术的准确性、通量和成本估计表明，它可以与目前基于高密度芯片的基因分型和基因表达分析相竞争。

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.