ELECTROCHEMICAL DETECTION USING FUNCTIONALIZED NANOPARTICLES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 该项目的总体目标是开发和测试新型光学传感器，以测量细胞间空间中生理相关分析物的交换。 这个特别的项目涉及测量细胞间离子活性，使用氢离子活性作为首选模型，使用功能化的纳米颗粒。我们将测试的功能化纳米颗粒包括与硅纳米颗粒结合的荧光分子（FITC）。原子力显微镜（AFM）显示FITC掺杂的硅纳米颗粒的直径小于100纳米。 初步研究表明，颗粒的内化发生在与细胞孵育的24小时内。 然而，细胞短期暴露于荧光颗粒改善了细胞外部的定位，并且时间过程实验表明细胞可以在不到30分钟内用颗粒标记。细胞短期暴露于纳米颗粒的另一个优点是降低毒性。细胞长期（10天）暴露于纳米颗粒似乎诱导细胞凋亡。我们打算使荧光传感器的数量多样化，以测量各种细胞外分析物的浓度，如钙，锌和氧。上述方法将作为更传统的生物电化学的替代技术，并提供新的见解细胞外化学结构域及其动力学。

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. The overall goal of this project involves the development and testing of novel optical sensors to measure exchange of physiologically relevant analytes in the intercellular space between cells. This particular project involves measurement of intercellular ionic activity, using hydrogen ionic activity as a model of first choice, using functionalized nanoparticles. The functionalized nanoparticles we will be testing include fluorescent molecules (FITC) conjugated to silicon nanoparticles. Atomic Force Microscopy (AFM) shows the FITC doped silicon nanoparticles are less than 100 nanometers in diameter. Preliminary studies indicate that internalization of particles occurs within 24 hours of incubation with cells. However, short term exposure of cells to fluorescent particles improves localization to the outside of cells, and time course experiments indicate that cells can be labeled with particles in less than 30 minutes. Another advantage to short term exposure of cells to nanoparticles is reduced toxicity. Long-term (10 days) exposure of cells to nanoparticles appears to induce apoptosis. We intend to diversify the number of fluorescent sensors to measure concentrations of a variety of extracellular analytes, such as calcium, zinc and oxygen. The approaches described above will serve as alternative techniques to more conventional bioelectrochemistry and provide new insights into extracellular chemical domains and their dynamics.