Localized Chemical Detection with Maintained Single Protein Channel Recordings on Nanoneedle Probes

在纳米针探针上保持单蛋白通道记录的局部化学检测

• 批准号: 2108368
• 负责人: Ryan White
• 金额: $ 40.5万
• 依托单位: University of Cincinnati Main Campus
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2108368&HistoricalAwards=false
• 关键词: Localized; Chemical; Detection; Maintained; Single

With the support of the Chemical Measurement and Imaging (CMI) Program in the Division of Chemistry, and partial funding from the Cellular Dynamics and Function Cluster in the Division of Molecular and Cellular Biosciences, Professor Ryan White of the University of Cincinnati Main Campus is studying new ways for measuring the release of small molecules at an interface with spatial resolution on the single biological cell and sub-cellular size scales. This measurement method is particularly significant in measuring cell signaling where small molecule messengers travel between cells dictating how they interact with their neighboring cells and environment. Because of the generality of the sensing method, the approach can be applied to a wide range of cell types, including non-neuronal cells in the brain. The proposed approach by Professor White and his team involves embedding a cellular membrane protein receptor, or protein channel, at the end of a nanoneedle electrode that can be placed in proximity to various interfaces for localized detection. In order the achieve this detection, the project will overcome the major challenge in protein channel measurements for analytical purposes – the ability to control and maintain a single channel through the duration of an experiment. The long-term goal of the research program is to develop probes that create “artificial synapses” to monitor the dynamics and heterogeneity of the cell membrane and cellular microenvironment. The work will provide broad impact in measurement science and new knowledge in the measure of cellular communication in the brain. This impact will reach a diverse group of students and scientists through the development of a program targeting the increased and sustained participation of first-generation college students in undergraduate research, through the creation of a strong community and culture.The overarching goal of this proposal is to develop nanoscale ion channel probes for imaging and localized molecular and ion sensing at interfaces with high spatial resolution (nanometer-micrometer). The impactful research will expand localized detection abilities to analytes that are not accessible with current methodologies, because the analytes are neither optically nor electrochemically active. This need is particularly significant in measuring cell signaling where messengers travel between cells, dictating how they interact with their environment. The measurements enabled by the new method will facilitate discoveries of how astrocytes, with contacts to the neurovascular system and neurons, differentially signal between the two which is critical for networked communication in the brain. Currently, however, there is a lack of generalizable tools that can measure molecules localized at a cell surface. The use of ion channel probes for scanning ion conductance microscopy (SICM), where protein channels are embedded in a lipid bilayer at the end of a probe, is a promising method to leverage ion channel activity for ion and molecular detection with the imaging capabilities of SICM. However, major hurdles exist that hinder their use. To overcome these challenges, Professor White and his team will develop a new SICM imaging mode that employs metal nanoneedle probes that support ion channel measurements with dramatically reduced probe size over existing approaches for localized detection at interfaces. They will demonstrate the ability to perform localized detection on test-bed inorganic substrates and the surface of astrocyte cells. These measurements are anticipated to enable the first direct, localized detection of adenosine triphosphate (ATP) release from astrocytes. The broader impacts of the work lie in the ability of the proposed measurement method to provide new knowledge about how biological systems (single cells, tissue slices) communicate via the release of small molecules. This impact will reach a diverse group of students and scientists through the development of a "Pathway to Undergraduate Research" program that targets increased and sustained participation of first-generation college students in undergraduate research through the creation of a strong community and culture.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

With the support of the Chemical Measurement and Imaging (CMI) Program in the Division of Chemistry, and partial funding from the Cellular Dynamics and Function Cluster in the Division of Molecular and Cellular Biosciences, Professor Ryan White of the University of Cincinnati Main Campus is studying new ways for measuring the release of small molecules at an interface with spatial resolution on the single biologic cell and sub-cellular size scales.这种测量方法在测量细胞信号传导方面特别重要，小分子使者在细胞之间传播，决定它们与邻近细胞和环境的相互作用。由于传感方法的普遍性，该方法可以应用于多种细胞类型，包括大脑中的非神经元细胞。怀特教授及其团队提出的方法涉及将细胞膜蛋白受体或蛋白质通道嵌入纳米电极的末端，该纳米德尔电极可以将其放置在各种接口以进行局部检测。为了实现这一检测的成就，该项目将克服蛋白质通道测量值的主要挑战，以进行分析目的 - 通过实验的持续时间控制和维持单个通道的能力。研究计划的长期目标是开发产生“人造突触”的问题，以监测细胞膜和细胞微环境的动态和异质性。这项工作将为测量科学和新知识提供广泛的影响，以测量大脑的细胞通信。通过制定一项计划，通过建立建立强大的社区和文化的计划，该计划将通过制定一项针对第一代大学学生参与本科研究的计划的计划来吸引一群学生和科学家。该提议的总体目标是开发纳米级离子渠道问题，以促进与较高的分子和离子感应的互动，并在较高的Spatial Sesigration（Nanansimential（Nanansimential）互动（Nannosial Spatial Sestrolution）（Nanansortor）（Nanansortor）（Nanansortor）中。有影响力的研究将将局部检测能力扩展到当前方法无法访问的分析物，因为分析物在光学上或电化学上是活跃的。这种需求在测量细胞之间在细胞之间传播的细胞信号传导，决定它们与环境之间的相互作用尤为重要。新方法实现的测量结果将有助于发现星形胶质细胞与神经血管系统和神经元的接触方式，这两者之间的差异信号对于大脑中的网络通信至关重要。但是，目前缺乏可概括的工具，可以测量在细胞表面定位的分子。在探针结束时将离子通道用于扫描离子电导显微镜（SICM），其中蛋白质通道被嵌入脂质双层中，是一种有望使用SICM成像功能的离子通道活性和分子检测的有望方法。但是，存在阻碍其使用的主要障碍。为了克服这些挑战，怀特教授和他的团队将开发一种新的SICM成像模式，该模式采用了金属纳米方问题，这些问题支持离子频道测量值，而在接口处的局部检测中，动态降低了探头大小。他们将证明对测试床的无机底物和星形胶质细胞表面进行局部检测的能力。预计这些测量值将使三磷酸腺苷（ATP）从星形胶质细胞释放。作品的更广泛的影响在于提出的测量方法提供有关生物系统（单细胞，组织切片）如何通过释放小分子传达的新知识的能力。这种影响将通过开发“通往本科研究的途径”计划来吸引一群学生和科学家，该计划通过建立强大的社区和文化来创建第一代大学学生从事本科研究的增加和持续参与。这奖反映了NSF的法规使命，并通过对基金会的知识优点进行评估，并以评估为众所周知的cristia cribia criitia criter critia criter criter critia criter criter criter critia criter criteria criter criteria criteria criperia criperia cr cr cr cr cordt cor而为支持的珍贵。

项目成果

Maintaining Single-Channel Recordings on a Silver Nanoneedle through Probe Design and Feedback Tip Positioning Control

• DOI: 10.1021/acs.jpcb.2c06275
• 发表时间: 2022-11-17
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY B
• 影响因子: 3.3
• 作者: Hussein，Essraa A.;White，Ryan J.
• 通讯作者: White，Ryan J.