HYBRID SENSORS & INTEGRATED PLATFORMS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 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，提供从单个细胞进行并行数据收集。这项工作包括混合传感器、结合电化学和光学方法的单个微电极以及集成平台的构建。 混合式传感器 自参考配置中使用的离子选择电极(SERIS探针)是一种高灵敏度、非侵入性的检测器，可以检测离子的缓慢扩散梯度，如来自上皮片或甚至来自单个细胞的质子或钙。然而，分泌的分子机制通常是从细胞内部控制的，这是传统的离子选择电极无法穿透的区域。为了同时获得有关离子分泌量和分泌控制机制的信息，正在建造一个双探针。这种双探针由一个离子选择电极和一个光纤光载体穿过尖端组成，用于将激发光传递到任意的报告分子。 在解决了几个技术问题后，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，必须对光纤进行硅烷化。硅烷化后，钙离子和质子离子选择性离子载体的选择性没有变化。要将激发光耦合到光纤中，需要在高(20 KHz)频率下振动光纤加载端，以确保均匀传输光学模式并减少激光散斑。该显微镜配备了基于荧光素的滤光片，以允许发射蓝色激光(Ar离子激光)和检测绿色发射光。 集成平台 当新技术与并行运行的现有技术相结合并提供相互佐证时，它们就更容易被接受。BRC已经维持了组合的电化学和电生理平台，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.