Research Initiation Award Grant: Ultrasensitive Plasmonic Nanoprobes for Monitoring of Intracellular Labile Iron Pools

本研究将开发一种微创的新传感方法，以解决在单细胞水平上理解细胞内不稳定铁池（LIP）的化学性质、分布和协作性的突出挑战。近年来，人们尝试测量细胞中的LIP；然而，LIP的精确测定仍然难以捉摸。用于LIP监测的超灵敏和潜在的细胞器特异性等离子体纳米探针的开发将提高对细胞内铁动力学和分布的理解，从而将对优化现有的螯合和补充铁疾病的治疗产生影响。这项研究还将为处于纳米技术研究前沿的非洲裔美国本科生和研究生提供研究培训机会。该项目将开发一种独特的方法，使用基于等离子体纳米结构的表面增强拉曼散射（SERS）来选择性地感知LIP，具有纳米摩尔灵敏度。这项工作涉及由银纳米晶体构建的二维超晶格的生物模板辅助纳米图案，这些纳米结构的物理化学性质将使用各种分析工具进行表征，如TEM， SEM， UV-Vis光谱，FT-IR和拉曼光谱。利用数百万个细胞的裂解物，对等离子体检测LIP的灵敏度和选择性进行优化和评估，该项目的最终目标是将最佳等离子体纳米结构整合到光纤的表面上，这样它就可以插入到单个细胞隔间中，并用作超灵敏的纳米探针，用于实时监测体外LIP。超灵敏的lip特异性等离子体纳米探针有望在阐明细胞内铁运输和动力学方面取得重大进展。

This study will develop a minimally invasive new sensing approach that addresses outstanding challenges of understanding the chemical nature, distribution and cooperativeness of the intracellular labile iron pool (LIP), at the single-cell level. In recent years, attempts have been made to measure the LIP in cells; however, the precise determination of the LIP remains elusive. The development of ultrasensitive and potentially organelle-specific plasmonic nanoprobes for LIP monitoring will improve the understanding of intracellular iron dynamics and distribution, and will thus have an impact on the optimization of existing chelation and supplementation therapies for iron disorders. The study will also provide research training opportunities for African American undergraduate and graduate students at the forefront of nanotechnology research. The project will develop a unique approach using plasmonic nanostructure-based surface-enhanced Raman scattering (SERS) for selective sensing of the LIP, with nanomolar sensitivity. The work involves biotemplate-assisted nanopatterning of two-dimensional superlattices constructed from silver nanocrystals, and the physicochemical properties of these nanostructures will be characterized using a variety of analytical tools such as TEM, SEM, UV-Vis spectrometry, FT-IR, and Raman spectroscopy. The sensitivity and selectivity for the plasmonic detection of the LIP will be optimized and evaluated using lysates from millions of cells, and the eventual goal of the project is to integrate the optimal plasmonic nanostructures onto the facet of an optical fiber, so it can be inserted to individual cellular compartments, and used as an ultrasensitive nanoprobe for real-time monitoring of the LIP in vitro. It is expected that considerable progress in the elucidation of intracellular iron trafficking and dynamics will be achieved by the development of ultrasensitive LIP-specific plasmonic nanoprobes.