Optical Encoding for Large Numbers of Samples

描述（由申请人提供）：描述了一种新的光学编码方法，用于标记大量样品，该方法基于测量含有不同比例稀土红外上转换荧光粉（RUP）标签的样品的发射比。初步数据表明，使用四色系统可以对bbb2.5 x 104样品进行多重处理。虽然以前用单个RUP材料标记单个样本的例子是已知的，但是基于单个报告粒子中多于一个RUP的比率的多路复用的使用极大地扩展了可以识别的单个样本的数量。稀土发射体在可见光区存在非重叠、非常窄的发射、在980 nm激发波长处缺乏样品自身荧光以及完全没有光漂白，使得高度复用成为可能。为了在试管中执行10,000个寡核苷酸微阵列实验，可以将感兴趣的10,000个寡核苷酸连接到10,000个不同的RUP样品上，并进行正常杂交。在一个能够分选8-1万个样品/秒的流式细胞仪内的两个独立的检测器上分析红/绿目标比率和RUP标签，可以在几分钟内分析数百万个基因表达产物。

DESCRIPTION (provided by applicant): A new optical encoding method for labeling large numbers of samples based on measurement of emission ratios in samples containing various ratios of rare earth infrared upconverting phosphor (RUP) labels is described. Preliminary data suggest it may be possible to multiplex >2.5 x 104 samples using a four colors system. While previous examples of labeling individual samples with a single RUP material are known, the use of multiplexing based on the ratios of more than one RUP within a single reporter particle vastly expands the number of individual samples that can be identified. The presence of non-overlapping, very narrow emissions in the visible region for the rare earth emitters, lack of sample autofluorescence at the 980 nm excitation wavelength and the total absence of photobleaching makes the high degree of multiplexing possible. To perform a 10,000 oligonucleotide microarray experiment in a test tube, the 10,000 oligonucleotides of interest could be attached to 10,000 different RUP samples and subjected to normal hybridization. Analysis of the red/green target ratio and the RUP label, on two separate detectors within a flow cytometer capable of sorting 8-10 thousand samples/second, could allow millions of gene expression products to be analyzed in a few minutes.