Multianalyte Nanoprobe for Neurochemicals

描述(申请人提供)：在对单个神经元和神经网络的细胞和细胞外研究中，监测电生理和神经化学活动是必不可少的。电化学探针现在已经被制造成适合这类研究的尺寸，但许多神经递质不能用电化学方法检测。此外，对化学敏感的电极经常消耗其目标分析物；在感兴趣的亚微米尺度上，这可能导致正在研究的系统的重大扰动。智能光学系统(IOS)与加州大学洛杉矶分校(UCLA)合作，提议为神经化学的细胞水平研究创造一种新工具--一种可以测量亚微米体积中多种分析物浓度的探测器。在这个建议的传感器中，一个多通道光波导结构，逐渐变小，尺寸小于它使用的光的波长，将使用与目标物质发生反向反应的荧光指示器来功能化。这一独特的探头将能够在毫秒级的时间尺度上连续监测局部的神经化学浓度。在拟议项目的第一阶段，内部监督办公室将建造3通道和4通道的“纳米探测器”，并与加州大学洛杉矶分校协商，用化学和生物化学为基础的识别系统激活它们，以识别感兴趣的分析物。光学激活的交联会将有机指示剂固定在可渗透的聚合物“点”中，直接位于光学通道的近场中，用于检测离子物种(例如，钙)和小分子。对于其他感兴趣的物种，将利用光活化结合在光学领域形成一层专利的生化“可逆化学识别单元”。在制造后，这些探头将在包含以下内容的库存溶液中进行校准 然后用于研究体外(培养的)神经元和脑片中的细胞外分析物水平，以证明它们在研究至关重要的神经生物学现象方面的有效性。在第二阶段，将研究体内应用。最终，iOS-UCLA团队计划将这些光学神经纳米探测器与类似大小的基于微电极的传感器结合起来，创造出具有大量(&gt；100)多功能探测器的阵列，用于在遥不可及的范围内同时绘制神经活动的电和化学图谱，以及目前最先进技术无法达到的化学细节水平。

DESCRIPTION (provided by applicant): In cellular and extracellular studies of individual neurons and neural networks, it is imperative to monitor both electrophysiological and neurochemical activity. Electrochemical probes have now been fabricated at a size scale appropriate for such studies, but many neurotransmitters cannot be detected electrochemically. Furthermore, chemically-sensitive electrodes often consume their target analytes; at the sub-micron size scales of interest, this can lead to significant perturbation of the system being studied. Intelligent Optical Systems (IOS), working with the University of California Los Angeles (UCLA), proposes to create a new tool for cell-level studies of neurochemistry - a probe that can measure the concentrations of multiple analytes in sub- micron volumes. In this proposed sensor a multi-channel optical waveguide structure, tapered to a size smaller than the wavelengths of light it uses, will be functionalized with fluorescent indicators that react reversily with target substances. This unique probe will enable continuous monitoring of localized neurochemical concentrations on a time scale of milliseconds. During Phase I of the proposed project, IOS will construct 3- and 4-channel "nanoprobes" and, consulting with UCLA, will activate them with chemical- and biochemical-based recognition systems for analytes of interest. Optically-activated crosslinking will immobilize organic indicators embedded in permeable polymer "dots" directly in the near field of optical channels for detection of ionic species (e.g., Ca++) and small molecules. For other species of interest, photoactivated binding will be used to form a layer of patented biochemical "reversible chemical recognition units" in the optical field. After fabrication, these probes will be calibrated in stock solutions containing their target molecules, and then used to study extracellular analyte levels in ex vivo (cultured) neurons and brain slices to demonstrate their effectiveness in studying critically important neurobiological phenomena. In Phase II, in vivo applications will be investigated. Ultimately, the IOS-UCLA team plans to combine these optical neuro-nanoprobes with microelectrode-based sensors of similar size to create arrays with large numbers (>100) of multifunction probes for simultaneous electrical and chemical mapping of neurological activity at a scale out of reach, and a level of chemical detail currently out of reach with state-of-the-art technology.