SBIR Phase I: Microfluidic Technology for Single Cell Multi-Omic Sample Preparation

小型企业创新研究(SBIR)第一阶段项目的更广泛影响/商业潜力是将一种能够颠覆现有样品制备市场的自动化微流控技术商业化。这个市场预计到2025年将超过53.2亿美元，这是由新的和可自动化的技术平台的发展推动的。这些技术平台不仅促进了精准医学的发展，也促进了我们对细胞行为背后的基本机制的理解。在这方面，自动化样品制备平台因其易用性和与上游细胞捕获技术(例如单细胞选择、稀有细胞浓缩)以及下游分析技术(例如DNA/RNA测序、质谱仪)的集成潜力而受到青睐。自动化平台不需要科学家复制由不同学术实验室开发的复杂的内部协议，而是允许用户只需加载样本就可以自由离开。然而，这一市场上现有的平台受到其基本单室设计的限制，使得复杂系统的处理具有极大的挑战性。这个小企业创新研究(SBIR)第一阶段项目旨在展示一种根本不同的方法来处理多组单细胞的可行性。而不是依赖单隔室设计(例如，液滴微流体、微孔技术、阀门和腔体微通道、管基成套件等)。在细胞处理方面，该项目将使用基于微柱的新型微流控技术，实现单细胞基因组的高效分离和分离。在这里，我们微流控通道内的微柱阵列可以在物理上将细胞裂解过程中的基因组DNA与蛋白质和RNA分离，其方式与现有的分析方法兼容。在第一阶段，我们将以我们的初步结果为基础，展示从同一细胞中分离出基因组和转录组，以及在亚硫酸氢盐治疗前后对基因组、转录组和甲基组进行三体分离之前和之后对基因组DNA进行扩增和测序的能力。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project is to commercialize an automatable microfluidic technology capable of disrupting the existing sample preparation market. This market, projected to surpass $5.32 billion USD by 2025, is driven by the development of new and automatable technology platforms. These technology platforms not only facilitate the development of precision medicine, but also our understanding of the fundamental mechanisms behind cellular behavior. Here, automated sample preparation platforms are favored for their ease of use and integration potential with both upstream cell capture technologies (e.g. single cell selection, rare cell enrichment) as well as downstream analysis technologies (e.g. DNA/RNA sequencing, mass spectrometry). Rather than requiring scientists to reproduce complex in-house protocols developed by different academic laboratories, automated platforms would allow users the freedom of simply loading a sample and walking away. However, existing platforms in this market are limited by their fundamental single-compartment design, making the processing of complex systems highly challenging.This Small Business Innovation Research (SBIR) Phase I project proposes to demonstrate the feasibility of a fundamentally different approach to multi-omic single cell processing. Instead of relying on a single-compartment design (e.g. droplet microfluidics, microwell technologies, valved and chambered microchannels, tube-based kits, etc.) for cell processing, this project will use novel micropillar-based microfluidic technology for high efficiency separation and isolation of single cell omes. Here, micropillar arrays within our microfluidic channels serve to physically size-separate genomic DNA from proteins and RNA during cell lysis in a manner compatible with existing analysis methods. In Phase I, we will build upon our preliminary results by demonstrating bi-omic isolation of the genome and transcriptome from the same cell along with the ability to amplify and sequence the genomic DNA before and after bisulfite treatment for tri-omic isolation of the genome, transcriptome, and methylome.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.