MRI: Development of an Instrument for Single Cell Electrical Stimulation and Analysis

了解最小的生物单位-单个细胞-如何对电刺激做出反应，对于开发用于组织修复的精确电刺激治疗，发明用于组织再生的新型导电生物材料，以及确定用于癌症治疗的新的敏感生物标志物至关重要。阿克伦大学的研究人员通过主要研究仪器(MRI)计划，致力于开发一种新的仪器，以发现和解释单个细胞对特定电刺激的特定反应。通过整合微流体和生物纳米技术的进步，该仪器将具有以下独特的能力和意义：a)精确地向单细胞提供受控的电刺激，b)收集和分析细胞分泌的多种类型的分子，c)直接评估产生的单细胞反应，而不受周围细胞、组织或物质的干扰信号，以及d)在刺激过程中“免接触”细胞操作，避免对细胞进行酶促物理处理。凭借这些独特的能力，该仪器将促进对细胞如何感知、功能、增殖和通信响应电刺激的基本理解，这无疑将对组织工程、再生医学、癌症生物学、生物材料和仿生设备产生革命性影响。该仪器将作为一个重要的研究设施，为阿克伦大学的教职员工和学生建立新的研究和培训能力，特别是该大学新成立的仿生研究和创新中心和综合生物科学博士项目，并使阿克伦-克利夫兰地区其他研究机构、行业和政府实验室的生物科学研究人员受益。该设计理念和仪器还将通过课程开发、阿克伦大学的“工程学术领域日益多样化”和“工程领域的女性”计划，为研究生、本科生和K-12学生提供极好的培训机会。为了推进当前的细胞研究，迫切需要破译和操纵单个细胞对可定义的电刺激的特定反应。然而，到目前为止，还没有一种通用的仪器可以精确地向单个细胞传递各种电刺激，收集和分析细胞分泌的分子。这个NSF项目旨在开发一种新的仪器，能够破解任何类型的单细胞的生物电码，建立新的研究能力，用于单细胞水平的操作和表征。为了达到预期的功能和规格，该仪器将由1)固定式表面声波(SSAW)细胞聚焦器组成，其中单个细胞将被聚焦在微通道的中心线上，2)蛇形微通道中的微电极阵列以连续的方式对单个细胞施加受控的电刺激，3)细胞培养室阵列用于培养受刺激的细胞，并形成目标分子-微粒聚集体，以及4)电阻脉冲传感器阵列用于测量聚集体和确定细胞分泌体中多种类型的分子浓度。最终，上述四个主要部分将整合为一个紧凑而强大的细胞研究新工具。新仪器将直接影响1)心脏修复和再生、2)神经组织工程、3)电刺激辅助伤口愈合和4)神经再生导电生物材料的先进研究。

Understanding how the smallest organism unit, a single cell, responds to electrical stimulations is vital for developing precision electrical stimulation treatments for tissue repair, inventing novel conductive biomaterials for tissue regeneration, and identifying new sensitive biomarkers for cancer therapy. Researchers at University of Akron through the Major Research Instrument (MRI) program are working on developing a new instrument to discover and interpret specific responses of single cells to defined electrical stimulations. By integrating the advances in microfluidics and bio-nanotechnology, the instrument will have the following unique capabilities and significances: a) precise delivery of controlled electrical stimulation to single cells, b) collection and analysis of multiple types of molecules secreted by cells, c) direct assessment of resultant single cell responses without the interfering signals from surrounding cells, tissues or substances, and d) "touch free" cell manipulation during the stimulation procedure, avoiding enzymatic, physical treatment of cells. With these unique capabilities, this instrument will advance basic understanding of how cells sense, function, proliferate, and communicate in response to electrical stimulations, which will undoubtedly have transformative impacts on tissue engineering, regenerative medicine, cancer biology, biomaterials and biomimetic devices. The instrument will serve as a major research facility that will build new research and training capacities for faculty members and students at University of Akron, especially for the university's newly created Biomimicry Research and Innovation Center and Integrated Bioscience Ph.D. Program, and benefit bioscience researchers from other research institutes, industry and government laboratories in Akron-Cleveland area. The design concept and the instrument will also provide excellent training opportunities for graduate, undergraduate and K-12 students via course development, University of Akron's "Increasing Diversity in Engineering Academics", and "Women in Engineering" programs. Deciphering and manipulation of specific responses of single cells to definable electrical stimulations is urgently needed to advance current cell research. However, to date a universal instrument is unavailable than can precisely deliver various electrical stimulation to individual cells, collect and analyze secreted molecules by cells. This NSF project aims to develop a new instrument that will enable cracking the bioelectrical code of single cells of any type, building new research capacities for single cell level manipulation and characterization. To achieve the desired functions and specifications, the instrument will consist of 1) a standing surface acoustic wave (SSAW) cell focuser in which single cells will be focused in the centerline of microchannels, 2) an array of microelectrodes in a serpentine microchannel to apply controlled electrical stimulations to single cells in a continuous flow, 3) an array of cell culture chambers to culture the stimulated cells, and form target molecule-microparticle aggregates, and 4) a resistive pulse sensor array to measure the aggregates and determine multiple type of molecule concentrations in cell secretome. Ultimately, the above four major parts will be integrated into a compact yet powerful new instrument for cell research. The new instrument will directly impact the advanced researches on 1) Cardiac repair and regeneration, 2) Neural tissue engineering, 3) Electrical stimulation assisted wound healing and 4) Conductive biomaterials for nerve regeneration.