High density array biosensors for spatial mapping of cellular gradients and flux

建议编号：1403582P.I.：Jenna L.Rickus标题：用于细胞梯度和通量空间映射的高密度阵列生物传感器细胞表面的小分子和离子的通量携带着重要的信息，这些信息在细胞内部和外部与相邻细胞交流。生物学中这种重要的细胞通量的例子包括神经元中神经递质的释放和再摄取，癌细胞中适应细胞新陈代谢的葡萄糖运输，以及植物发育期间重力感应中的离子通量。目前还没有一种方法可以量化芯片形式的许多单个细胞的真实细胞通量和生物分子梯度的空间地图和动力学。该项目将应用纳米/微电子学、数学建模和生物传感器方面的最新进展和专业知识来创造这样的技术。这项工作的结果将是一种芯片平台技术，可以在广泛的细胞系统中复制，以回答有关细胞流量的基本生物学问题，这些问题目前无法回答。例如，一个新切除的肿瘤可以被快速分析，以检查候选药物如何影响不同细胞群体的葡萄糖代谢，或者神经递质动态可以与电活动信息相结合，以更好地了解学习和记忆过程中神经元网络的信息处理。细胞表面小分子和离子的通量和由此产生的浓度梯度携带着重要的信息，这些信息在细胞内部和外部与邻近细胞交流。自参照生物传感器是目前量化细胞通量动态的最佳方法，但该方法仅限于一次在一个细胞上的一个空间位置进行测量。此外，这种方法不便于携带，不能用于诸如空间飞行任务、环境监测或高通量生物医学应用等偏远地点的传感。目前还没有一种方法可以量化芯片形式的单个细胞的真实细胞通量和生物分子梯度的空间图谱和动力学。这项工作的总体目标是通过创建可单独寻址的纳米/微米级电化学传感器的一维和二维阵列，创建一种在单细胞水平上绘制生物分子梯度和通量的空间和时间图谱的技术。该提案开发并整合了新的解决方案，以解决创建这种设备的技术挑战，包括克服噪声限制的同步检测，减少传感器和细胞之间精确对准需求的现场可编程方法，实现生物识别层和多分析物传感的高空间分辨率的电极可寻址生物功能化，以及考虑生物信号和传感器在空间和时间上的功能来设计和优化电极阵列的数学框架。重要的是，这些技术进步中的每一项都具有超越通量传感的应用，并更广泛地应用于纳米生物传感领域。这项工作的最终结果将是一种可在广泛的蜂窝系统中复制的芯片平台技术。该奖项由两个项目联合颁发--(1)化学、生物工程、环境和运输系统司(工程局)的纳米生物传感，以及(2)生物基础设施部(生物科学局)的生物研究仪器开发。

Proposal Number: 1403582P.I.: Jenna L. RickusTitle: High density array biosensors for spatial mapping of cellular gradients and fluxThe flux of small molecules and ions at the surface of cells carries important information that is communicated both internally to the cell and externally to neighboring cells. Examples of such important cellular flux in biology include neurotransmitter release and reuptake in neurons, glucose transport for adapted cell metabolism in cancer cells, and ion flux in plant gravity sensing during development. No method currently exists that can quantify the spatial map and dynamics of true cellular flux and biomolecule gradients from many single cells in a chip format. This project will apply recent advances and expertise in nano/microelectronics, mathematical modeling, and biosensors to create such a technology. The outcome of the work will be an on chip platform technology that can be replicated for a wide range of cellular systems to answer basic biological questions regarding cellular flux that cannot be answered today. For example, a freshly resected tumor could be rapidly analyzed to examine how a drug candidate affects glucose metabolism of the heterogeneous cell population or neurotransmitter dynamics could be integrated with electrical activity information to better understand information processing of neuronal networks during learning and memory.The flux and resulting concentration gradients of small molecules and ions at the surface of cells carry important information that is communicated both internally to the cell and externally to neighboring cells. Self-referencing biosensors are currently the best method for quantifying cellular flux dynamics, but this method is limited to measurements at one location in space on one cell at a time. In addition, this method is not portable and cannot be used for sensing in remote locations such as on space missions, in environmental monitoring, or in high throughput biomedical applications. No method currently exists that can quantify the spatial map and dynamics of true cellular flux and biomolecule gradients from single cells in a chip format. The overall objective of this work is to create a technology for the spatial and temporal mapping of biomolecule gradients and flux at the level of single cells by creating 1D and 2D arrays of individually addressable nano/microscale electrochemical sensors. The proposal develops and integrates novel solutions to the technical challenges of creating such a device including synchronous detection to overcome noise limitations, a field-programmable approach to reduce the need for precision alignment between sensors and cells, electrode addressable biofunctionalization to achieve high spatial resolution of bio-recognition layers and multi-analyte sensing, and a mathematical framework to design and optimize electrode arrays considering both the biological signal and sensor function in space and time. Importantly each of these technical advances has application beyond flux sensing and applies more broadly to the field of nano-biosensing. The end result of the work will be an on-chip platform technology that can be replicated for a wide range of cellular systems.This award is being made jointly by two Programs- (1) Nano-Biosensing, in the Division of Chemical, Bioengineering, Environmental and Transport Systems (Engineering Directorate), and (2) Instrument Development for Biological Research, in the Division of Biological Infrastructure (Biological Sciences Directorate).