Pregibon Daniel C, Toner Mehmet, Doyle Patrick S
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Science. 2007 Mar 9;315(5817):1393-6. doi: 10.1126/science.1134929.
High-throughput screening for genetic analysis, combinatorial chemistry, and clinical diagnostics benefits from multiplexing, which allows for the simultaneous assay of several analytes but necessitates an encoding scheme for molecular identification. Current approaches for multiplexed analysis involve complicated or expensive processes for encoding, functionalizing, or decoding active substrates (particles or surfaces) and often yield a very limited number of analyte-specific codes. We present a method based on continuous-flow lithography that combines particle synthesis and encoding and probe incorporation into a single process to generate multifunctional particles bearing over a million unique codes. By using such particles, we demonstrate a multiplexed, single-fluorescence detection of DNA oligomers with encoded particle libraries that can be scanned rapidly in a flow-through microfluidic channel. Furthermore, we demonstrate with high specificity the same multiplexed detection using individual multiprobe particles.
高通量筛选用于基因分析、组合化学和临床诊断,受益于多重分析,它允许同时检测几种分析物,但需要一种用于分子识别的编码方案。当前的多重分析方法涉及用于编码、功能化或解码活性底物(颗粒或表面)的复杂或昂贵过程,并且通常产生非常有限数量的分析物特异性代码。我们提出了一种基于连续流光刻的方法,该方法将颗粒合成、编码和探针掺入结合到单个过程中,以生成带有超过一百万个唯一代码的多功能颗粒。通过使用此类颗粒,我们展示了使用编码颗粒文库对DNA寡聚物进行多重单荧光检测,该文库可在流通微流体通道中快速扫描。此外,我们使用单个多探针颗粒以高特异性证明了相同的多重检测。