High Density 3D Printed Microfluidics With Open Source Resins for Biomedical Applications

项目摘要 微流控(单芯片实验室)是一项非常有前途的技术，适用于非常广泛的生物医学领域。 应用包括药物发现；组织工程；医疗诊断；癌症筛查 基于罕见的细胞检测、蛋白质、DNA或微型RNA生物标记物，以及最近的循环 外显体。这项提议旨在通过开发3D技术来彻底改变生物医学微流控生态系统 打印以常规地创建用于生物医学的非常小的、密集集成的微流控设备 科学。利用当前的微流控制造技术，这种器件是不可能的，通常 依赖于几个单独制造的层的仔细对准和粘合，每个层都有一个 二维零部件布局。相比之下，3D打印允许设备体积的所有3个维度都完全 用于元件布局和通道布线，为密集元件提供机会 集成度高，器件体积小(~5 mm~3)。例如，我们包括了一个初步的设备设计 一种基于多重细胞的检测方法，可同时检测基因型(相关基因的存在)和表型 (电池特性和/或行为)在仅2.2 mm×2.2 mm×1 mm的器件体积中。该设备 包括细胞生长室，用于mRNA捕获和荧光测量的整体，以及 一体式泵和阀。此外，印刷运行1小时可实现快速制造和测试周期 极大地加快了设备开发。这项提议旨在开启一种良性循环，在这种良性循环中，3D 印刷微流体成为生物医学创新的颠覆性工具，这应该有一个 对人类健康有重大影响。 到目前为止，3D打印的关键制约因素是商用3D打印机和 用于制造构成微流体结构的必要微孔的树脂。我们的小组最近 演示了光敏树脂的正确配方与基础知识相结合 聚合光物理使低成本商用立体平版(SL)3D打印机能够 制造最小尺寸为100微米的空隙。本提案的目标1和2将 在这一成功的基础上，开发3D打印机和相应的低成本树脂来制造阀门 比我们已经演示的尺寸小40倍，以及具有横截面尺寸的流道 降至24 x 30微米。这些进展将在目标3中用于构建高密度微流控 探测(1)活细胞中多个基因的表达和(2)量化这些基因的活性的装置 单个设备中的电池。这些研究的总体目标是开发3D打印微流控 具有能够实现基于细胞的微型生物分析的特征尺寸的系统。

Project Summary Microfluidics (lab-on-a-chip) is a promising technology for an extremely broad range of biomedical applications including drug discovery; tissue engineering; point-of-care diagnostics; cancer screening based on rare cell detection, protein, DNA, or micro-RNA biomarkers, and more recently, circulating exosomes. This proposal aims to revolutionize the biomedical microfluidic ecosystem by developing 3D printing to routinely create very small, densely integrated microfluidic devices for the biomedical sciences. Such devices are not possible with current microfluidic fabrication techniques, which typically rely on careful alignment and bonding of a handful of individually fabricated layers, each of which has a 2D component layout. In contrast, 3D printing permits all 3 dimensions of the device volume to be fully utilized for component placement and channel routing, offering the opportunity for dense component integration and small device volume (~5 mm3). For example, we include a preliminary device design for a multiplexed cell-based assay that tests both genotype (presence of the relevant gene) and phenotype (cell characteristics and/or behavior) in a device volume of only 2.2 mm × 2.2 mm × 1 mm. The device includes cell growth chambers, monoliths for mRNA capture and fluorescence measurement, and an integrated pump and valves. Moreover, print runs <1 hour enable fast fabrication and test cycles to dramatically speed device development. This proposal intends to initiate a virtuous circle in which 3D printed microfluidics becomes a disruptive tool for biomedical innovation, which should have a substantial impact on human health. To date, the key inhibiting factor for 3D printing has been the inability of commercial 3D printers and resins to fabricate the requisite microvoids that comprise microfluidic structures. Our group recently demonstrated that proper formulation of photosensitive resins coupled with fundamental understanding of the polymerization photophysics enables a low cost commercial stereolithographic (SL) 3D printer to fabricate voids with minimum dimensions on the order of 100 µm. Aims 1 and 2 of this proposal will build on this success by developing a 3D printer and commensurate low cost resins to fabricate valves 40x smaller than what we have already demonstrated, and flow channels with cross section dimensions down to 24 x 30 µm2. These advances will be used in Aim 3 to construct high density microfluidic devices that probe (1) expression of multiple genes in live cells and (2) quantify the viability of those cells in a single device. The overall objective of these studies is to develop 3D printed microfluidic systems with feature dimensions that are enabling for miniaturized cell-based bioanalysis.