Single molecule DNA/RNA transport and Raman scattering readout in a coupled nanochannel/nanopore sequencing system.

耦合的单分子DNA/RNA输运和拉曼散射读数 纳米通道/纳米孔测序系统 Armonica Technologies，Inc.提议开发一种新型的、高通量、无标签、高度 基于廉价的纳米级图案化和自适应的精确、长读DNA测序平台 集合。平台由纳米通道(横截面尺寸为~100 nm)组成；曲折的 由胶体纳米颗粒自组装形成的(卷曲的、3D的)纳米孔；纳米颗粒屏障 跨纳米沟道放置；以及金属-绝缘体-金属(MIM)场增强结构 纳米沟槽屋顶。在操作中，单链或双链DNA被部分拉伸成 纳米通道中的线性构型被结合到通道中的势垒阻挡并强制 (通过电场)通过屋顶上曲折的纳米孔转移。屋顶上的MIM结构 局部增强外加激光光源的电磁场，允许表面增强相干 反斯托克斯拉曼散射(SECARS)检测单个碱基通过 电磁热点，从而提供单碱基灵敏度和空间定位。独一无二的 单个碱基的拉曼光谱允许无标记测序。光学检测可以大量地 并行操作，因为唯一的要求是通过多于一个光学元件来分离毛孔 波长，这在制造过程中很容易实现。该平台的一个重要特点是 多孔的屋顶允许引入寡核苷酸、小蛋白和DNA结合/DNA 加工酶，允许对纳米通道中的DNA进行可选的操作和修饰。 这个第一阶段项目的目标是：优化MIM结构，以实现单基座的灵敏度和空间 本地化；展示单碱基敏感性

Single molecule DNA/RNA transport and Raman Scattering readout in a coupled nanochannel/nanopore sequencing system Armonica Technologies, Inc. is proposing to develop a novel, high-throughput, label-free, highly accurate, long-read DNA sequencing platform based on inexpensive nanoscale patterning and self- assembly. The platform consists of nanochannels (cross section dimensions of ~ 100 nm); tortuous (convoluted, 3D) nanopores formed by self-assembly of colloidal nanoparticles; nanoparticle barriers placed across the nanochannels; and a metal-insulator-metal (MIM) field enhancement structure atop the nanochannel roof. In operation, single- stranded- or double-stranded-DNA is partially stretched into a linear configuration in the nanochannels, is blocked at barriers incorporated into the channels and forced (by electric field) to translocate through the tortuous nanopores in the roof. The MIM structure on the roof locally enhances the electromagnetic fields of applied laser sources allowing surface enhanced coherent anti-Stokes Raman scattering (SECARS) detection of individual bases as they pass through the electromagnetic hot spots, thus providing single base sensitivity and spatial localization. The distinct Raman spectra of the individual bases allow label-free sequencing. Optical detection allows massively parallel operation since the only requirement is separation of the pores by more than an optical wavelength, which is easily accomplished in the fabrication. An important feature of the platform is that the porous roofs allow introduction of oligonucleotides, small proteins, and DNA-binding/DNA processing enzymes, permitting optional manipulation and modification of the DNA in the nanochannels. The goals of this Phase I project are: to optimize the MIM structure for single base sensitivity and spatial localization; to demonstrate single base sensitivity