Microfluidics to explore ultrafast cell deformations to deliver large cargo via convective transport

项目摘要 将分子和颗粒递送至细胞对于增加分子生物学的科学理解是重要的。 过程和网络，检测诊断靶标中的细胞内生物标志物，以及基因修饰， 治疗细胞与当前交付方法相关的挑战包括限制 可以递送的分子（尤其是质粒DNA）、对靶细胞的损伤或修饰，以及对 小区处理吞吐量。在研究开发新的方法，可用于提供分子和 粒子更广泛地应用于许多细胞类型，发现了一种新的细胞行为， 以超过一毫秒的时间尺度迅速压缩。由于细胞受到快速挤压， 通过体积的暂时变化来响应，这导致压力驱动流过细胞膜， 恢复细胞体积，并作为副产物通过对流膜将细胞外试剂带入细胞。 现象本研究的目的是了解如何优化设备，利用新的生物物理 细胞压缩方案，其中快速时标（<1毫秒）、高应变（>30%）冲击细胞。这些 由于在芯片实验室中的应用，细胞的物理影响越来越重要， 分选和细胞工程。其次，将对微流控技术进行优化和测试， 探针和标记的递送，以及用于多种重要细胞类型的大转基因的转染。 在一个未探索的时间和幅度的区域内了解细胞机械反应可以使 新的方法来标记和重新编程细胞，这将是有效的广泛的细胞类型， 试剂

Project Summary The delivery of molecules and particles to cells is important for increased scientific understanding of molecular processes and networks, detecting intracellular biomarkers in diagnostic targets, and genetic modification of therapeutic cells. The challenges associated with current delivery approaches include limits to the size of molecules that can be delivered (especially plasmid DNA), damage or modification to target cells, and a limit to the cell processing throughput. In studies to develop new methods that can be used to deliver molecules and particles more broadly to many cell types, a novel cellular behavior was discovered that occurs as cells are rapidly compressed at timescales faster than a millisecond. As a result of fast compressions upon cells, cells respond by a temporary change of volume, which results in a pressure driven flow across the cell membrane to restore cell volume, and as a byproduct carries extracellular reagents into a cell through a convective phenomenon. The goal of this study is to understand how to optimize devices exploiting a new biophysical regime of cell compression in which fast timescales (<1 millisecond), high strain (>30%) to impact cells. These physical impacts of cells are increasingly important to understand due to applications in lab on a chip, cell sorting, and cell engineering. Secondly, the microfluidic technology will be optimized and tested for microfluidic delivery of probes and labels, as well as transfection of large transgenes for a variety of important cell types. The understanding of cell mechanical responses in an unexplored region of time and magnitude could enable new approaches to label and reprogram the cell that will be efficient to a broad range of cell types and reagents.