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，用于基于包含不同比例的稀土红外上转换荧光粉(RUP)标签的样品中的发射比的测量来标记大量样品。初步数据表明，使用四色系统对2.5x104个样品进行多色处理是可能的。虽然以前使用单个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。稀土发光体在可见光区存在不重叠的、非常窄的发射，在980 nm激发波长缺乏样品的自荧光，以及完全没有光漂白，使得高度的多路复用成为可能。为了在试管中进行10,000个寡核苷酸微阵列实验，可以将10,000个感兴趣的寡核苷酸连接到10,000个不同的RUP样本上，并进行正常杂交。在能够每秒分选8-10,000个样本的流式细胞仪中，在两个独立的检测器上分析红/绿目标比率和RUP标记，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.