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

提案编号：1403582 P.I.：珍娜湖Rickus职务：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重要信息，这些信息在细胞内部和外部与相邻细胞进行通信。生物学中这种重要的细胞通量的例子包括神经元中的神经递质释放和再摄取，癌细胞中适应细胞代谢的葡萄糖转运，以及植物发育期间重力感应中的离子通量。 目前还不存在可以以芯片形式量化来自许多单细胞的真实细胞通量和生物分子梯度的空间图和动力学的方法。 该项目将应用纳米/微电子学，数学建模和生物传感器的最新进展和专业知识来创建这样的技术。 这项工作的成果将是一种芯片平台技术，可以复制用于广泛的细胞系统，以回答今天无法回答的关于细胞通量的基本生物学问题。 比如说，新切除的肿瘤可以被快速分析以检查候选药物如何影响异质细胞群的葡萄糖代谢，或者神经递质动力学可以与电活动信息整合以更好地理解学习和记忆期间神经元网络的信息处理。细胞表面的小分子和离子的通量和由此产生的浓度梯度携带重要信息，既在内部与细胞连通，又在外部与相邻细胞连通。自参考生物传感器是目前量化细胞通量动态的最佳方法，但这种方法仅限于一次在一个细胞上的空间中的一个位置处进行测量。此外，这种方法不是便携式的，并且不能用于在远程位置进行感测，例如在空间任务中、在环境监测中或在高通量生物医学应用中。 目前不存在可以以芯片形式量化来自单细胞的真实细胞通量和生物分子梯度的空间图和动力学的方法。 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1D和2D阵列，在单细胞水平上创建用于生物分子梯度和通量的空间和时间映射的技术。 该提案开发并整合了针对创建这种设备的技术挑战的新解决方案，包括同步检测以克服噪声限制，现场可编程方法以减少传感器和细胞之间精确对准的需求，电极可寻址生物功能化以实现生物识别层的高空间分辨率和多分析物感测，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个都具有超出通量传感的应用，并且更广泛地应用于纳米生物传感领域。这项工作的最终成果将是一个芯片平台技术，可以复制到广泛的细胞系统。这个奖项是由两个程序联合作出-（1）纳米生物传感，在化学，生物工程，环境和运输系统司（工程理事会），和（2）生物基础设施司（生物科学理事会）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).