HYBRID SENSORS & INTEGRATED PLATFORMS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 作为BRC作为综合技术资源的一部分，我们努力将不同的实验方法结合在一起，提供来自单细胞的并行数据收集。 这项工作包括混合传感器，结合电化学和光学方法的单微电极，以及集成平台的构建。 混合传感器 自参考配置中使用的离子选择性电极（SERIS探针）是高灵敏度的非侵入性检测器，可检测来自上皮层甚至单细胞的离子（如质子或钙）的缓慢扩散梯度。 然而，分泌的分子机制通常是从细胞内控制的，这是常规离子选择性电极无法穿透的区域。 为了同时获得关于离子分泌量和控制分泌的机制的信息，正在构建双探针。 双探针由一个离子选择性电极与光纤光载体插入通过尖端，用于传递激发光到任意的报告分子。 在解决了几个技术问题之后，已经建造了一个原型探测器。 为了避免破坏离子载体的选择性，光纤必须进行硅烷化。 硅烷化后，离子载体的选择性不变，测量与钙和质子选择性离子载体。 将激发光耦合到光纤中需要以高（20 kHz）频率振动光纤加载端，以确保光学模式的均匀传递并减少激光散斑。 显微镜配备有基于荧光素的滤光器，以允许递送蓝色激光（氩离子激光）并检测绿色发射光。 综合平台 新技术如果与并行运作的现有技术相结合并相互印证，就会得到更普遍的接受。 BRC已经保持了电化学和电生理学平台的组合，后者包括单个微电极，电压，电流和膜片钳。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. As part of the BRC's role as an integrated technology resource, we strive to bring together different experimental approaches, providing parallel data collection from single cells. This effort encompasses hybrid sensors, single microelectrodes combining electrochemical and optic approaches, as well as the construction of integrated platforms. Hybrid Sensors Ion-selective electrodes used in the self-referencing configuration (SERIS probes) are highly sensitive, noninvasive detectors of slow diffusion gradients of ions such as protons or calcium from epithelial sheets or even from single cells. The molecular mechanisms of secretion, however, are usually controlled from within the cell, a domain into which a conventional ion-selective electrode cannot penetrate. To gain simultaneous information about the amount of ion secretion and the mechanisms controlling secretion, a dual probe is under construction. The dual probe consists of an ion-selective electrode with a fiber-optic light carrier inserted through the tip, for delivery of excitation light to an arbitrary reporter molecule. A prototype probe has been constructed, following resolution of several technical issues. To avoid destroying the selectivity of the ionophore, the optical fiber had to be silanized. After silanization, selectivity of the ionophore was unchanged, as measured with both calcium- and proton-selective ionophores. Coupling of excitation light into the optical fiber required vibrating the fiber loading end at high (20 kHz) frequencies, to ensure even delivery of optical modes and reduce laser speckle. The microscope was equipped with fluorescein-based filters to allow delivery of blue laser light (argon ion laser) and detection of green emission light. Integrated Platforms New technologies are more generally accepted when they are combined with existing techniques operating in parallel and provide mutual corroboration. The BRC already maintains combined electrochemical and electrophysiological platforms, the later encompassing single microelectrode, voltage, current and patch clamp.