A high throughput platform for rapid single cell surface mapping

用于快速单细胞表面绘图的高通量平台

基本信息

批准号：
1905786
负责人：
Jiang Zhe
金额：
$ 37.34万
依托单位：
University of Akron
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1905786&HistoricalAwards=false
关键词：
throughput platform rapid single cell

项目摘要

Cell surface charge, net electricity on cell surface, is a promising new biomarker for rare cell detection, cell sorting, and pathology stage monitoring of malignant cell. Cell surface charge pattern also reflects the in situ dynamic status of cell membrane, which is critical for membrane-regulated cell functions such as endocytosis, muscle cell contraction, nutrient transport, T cell activation and insulin release. Recent evidences show that a variety of cell functions including cell aggregation, cell division and cell migration can be modulated by manipulating cell surface charge patterns. Therefore rapid mapping of single cell surface charge patterns will provide a unique and important approach in advancing analysis of cells and regulation of their functions. However, to date rapid mapping of living cell surface charge remains a large challenge owing to lack of robust techniques capable of measurements at the micro and nano scale. Scanning ion conductance microscopy has been used for quantitative cell surface charge mapping. This method employs a single nano pipette to approach the cell surface point by point, resulting in a low efficiency, labor intensive and destructive measurement. The proposed research aims to address this long standing challenge by exploring a universal method for rapid, non-destructive characterization and mapping of cell surface charge. In addition, with educational and outreach activities, the proposed research is poised to enhance the interdisciplinary learning experience for undergraduate, graduate and K-12 students. The objective of this project is to explore a lab-on-a- chip platform for rapid, non-destructive living cell surface charge mapping. Such a device will 1) foster the breakthrough of discovering new biomarkers for cell identification, sorting and real time monitoring, and 2) greatly advance the knowledge in understanding the roles cell electrical properties play on cell functions. To achieve the objective, the following tasks will be pursued: 1) demonstration of acoustic cell manipulator that will rapidly focus/release individual living cell onto/off the probe surface in a continuous flow, 2) study of an array of nanopores that can accurately map the cell surface charge with high temporal resolution and accuracy, 3) implementation of signal multiplexing that will enable simultaneous measurement of surface charge distributions via a nanopore array, and 4) validation of the platform for rapid cell surface charge mapping using human dermal fibroblasts cells. The use of non-intrusive cell manipulation enables quick focus/release cells onto/off the nanopore array's surface, allowing rapid, high-throughput surface charge mapping in continuous flow. The use of nanopore array in combination with signal multiplexing enables rapid mapping of the cell surface charge characteristics at one time without complex measurement electronics.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.

细胞表面电荷，细胞表面上的净电力，是一种有希望的新生物标志物，用于稀有细胞检测，细胞分选和病理学阶段监测恶性细胞。细胞表面电荷模式还反映了细胞膜的原位动态状态，这对于膜调节的细胞功能至关重要，例如内吞作用，肌肉细胞收缩，营养转运，T细胞激活和胰岛素释放。最近的证据表明，可以通过操纵细胞表面电荷模式来调节多种细胞功能，包括细胞聚集，细胞分裂和细胞迁移。因此，单细胞表面电荷模式的快速映射将为细胞分析及其功能调节提供独特而重要的方法。然而，迄今为止，由于缺乏能够在微型和纳米尺度上进行测量的强大技术，活细胞表面电荷的快速映射仍然是一个巨大的挑战。扫描离子电导显微镜已用于定量细胞表面电荷映射。该方法采用单个纳米移液管来逐点接近细胞表面点，从而导致低效率，劳动密集型和破坏性的测量。拟议的研究旨在通过探索一种通用方法来解决这一长期存在的挑战，以快速，非破坏性表征和细胞表面电荷的映射。此外，借助教育和外展活动，拟议的研究有望增强本科，研究生和K-12学生的跨学科学习经验。该项目的目的是探索一个实验室芯片平台，用于快速，无损的活细胞表面电荷映射。这样的设备将1）促进发现用于细胞识别，分类和实时监测的新生物标志物的突破，以及2）极大地促进了了解细胞电特性在细胞功能上起着作用的知识。 To achieve the objective, the following tasks will be pursued: 1) demonstration of acoustic cell manipulator that will rapidly focus/release individual living cell onto/off the probe surface in a continuous flow, 2) study of an array of nanopores that can accurately map the cell surface charge with high temporal resolution and accuracy, 3) implementation of signal multiplexing that will enable simultaneous measurement of surface charge distributions via a nanopore array, and 4)使用人真皮成纤维细胞的快速细胞表面电荷映射的平台验证。非侵入性细胞操作的使用可以使焦点/释放单元在纳米孔阵列的表面上快速/释放细胞，从而可以在连续流中快速，高通量表面电荷映射。纳米孔阵列与信号多路复用结合使用，可以一次无需复杂的测量电子设备即可快速映射细胞表面电荷特性。该奖项反映了NSF的法定任务，并被认为值得通过基金会的知识分子优点和更广泛的审查标准通过评估来进行评估。