SBIR Phase I: A Nanofluidic Instrument for High-throughput Single-molecule Analysis

这个小企业创新研究（SBIR）第一阶段项目旨在开发一种商用纳米流控芯片，用于快速识别单个分子上的表观遗传标记。通过消除当前技术的两个主要限制，该仪器将极大地改进现有的表观基因组分析方法。所提出的设备将能够同时使用微小的染色质输入和高通量单分子观察自动量化多个表观遗传标记。该方法包括测量特异性结合到单个染色质片段上的表观遗传标记的抗体的不同荧光信号，这些片段通过纳米级流体通道内聚焦的激光进行电泳运输。本提案的目标是通过使用平行流体通道增加吞吐量并转换为自由空间光学系统，将实验室装置转变为原型商业产品。预计第一阶段的建议将导致包含96个平行纳米流体通道的器件的制造和自由空间光学系统的设计，这将促进这种转变。该项目更广泛的影响/商业潜力在于，它将产生商业化的产品，克服当前表观基因组技术的两个关键限制。目前的技术需要大量的输入材料，一次只能查询一个表观遗传标记。所提出的单分子分析方法可以克服这两个限制。预计这一努力将产生一种新的颠覆性表观基因组学技术，以服务于商业、学术和临床需求。通过开发一种自动化仪器，可以同时询问从非常小的细胞输入中提取的单个染色质分子上的多个表观遗传标记，该技术将使表观基因组分析比目前可能的信息丰富得多，成本更低。

This Small Business Innovation Research (SBIR) Phase I project aims to develop a commercial nanofluidic chip for the rapid identification of epigenetic marks on individual molecules. The instrument will dramatically improve existing methods of epigenomic analysis by removing two major limitations of the current technology. The proposed device will enable automated quantification of multiple epigenetic marks simultaneously using minute inputs of chromatin and high throughput single molecule observations. The approach involves measuring distinct fluorescent signals from antibodies bound specifically to epigenetic marks on individual chromatin fragments that are electrophoretically transported through lasers focused within a nanoscale fluidic channel. The objectives of this proposal are to transform this laboratory setup into a prototype commercial product by increasing the throughput using parallel fluidic channels and transforming to a free space optical system. It is anticipated that this Phase I proposal will result in the fabrication of devices containing 96 parallel nanofluidic channels and the design of the free space optical system that will facilitate this transformation.The broader impact/commercial potential of this project is that it will result in commercially available products that overcome two key limitations of current epigenomic technology. Current technology requires an abundant amount of input material and can query only one epigenetic mark at a time. The proposed single molecule analytical methods can overcome both of these limitations. It is anticipated that this effort will yield a new disruptive epigenomics technology to serve commercial, academic, and clinical needs. By developing an automated instrument that can interrogate multiple epigenetic marks simultaneously on single chromatin molecules extracted from very small inputs of cells, this technology will enable epigenomic analyses that are far more information rich and lower in cost than is currently possible.