Development of a Novel Multi-functional Single Cell Analyzer

描述(申请人提供)：细胞内各种参数的体外定量测量和分析正在成为细胞研究的一个重要方面。全面了解单个细胞对其生物环境和刺激的响应正在成为许多生物医学研究领域的基础，包括药物开发、纳米毒性研究、生物标志物发现、癌症诊断和治疗等许多领域。然而，目前还没有这样的仪器可以同时对细胞内的各种关键参数进行全面、定量的测量。 该项目旨在开发一种新型的多功能单细胞分析仪，该分析仪集成了体外测量许多重要细胞内参数的能力。新的细胞分析仪将解决生物医学研究中的一些关键问题，如细胞坏死和凋亡、干细胞分化和癌细胞生物学。该项目设计了三个具体目标，包括：1)开发锥形光纤微探头和相关仪器，用于原位测量细胞内的pH、温度和拉曼光谱；2)将锥形光纤微传感器与形态和结构监测能力相结合，构建单细胞分析仪；以及3)以纳米材料的细胞毒性为应用模型，对多功能单细胞分析仪进行评估和演示。 拟议的新型单细胞分析仪独一无二地得到了该项目中嵌入的许多关键创新的支持。免组装的微纳传感器探头将设计和制造在光纤锥体的远端，可以直接插入Livin细胞，以高分辨率原位定量测量各种重要参数。这些微型传感器与显微成像工具的创新集成将允许对细胞对生物环境和刺激的反应进行全面、系统的研究。以纳米粒子的细胞毒性为例，评价和演示单细胞分析仪的基本功能，也将为进一步的性能改进、系统优化和应用范围的扩大提供有价值的信息。 这项拟议的研究将由密苏里科技大学(密苏里S&T)的三名教授组成的多学科团队进行。这个项目所需的所有主要设备都在密苏里州S&T校区。团队成员在生物化学/生物分析、纳米材料的细胞毒性、光学微/纳米传感器和仪器以及分析仪器设计和实施方面拥有互补的技能。生命科学、电气工程和化学的跨学科项目将促进不同领域的教职员工和学生之间的强大合作，并加强密苏里州S&T的生物医学相关研究和基础设施。

DESCRIPTION (provided by applicant): In vitro quantitative measurements and analyses of various intracellular parameters is becoming an important aspect in cell research. Comprehensive understanding of a single cell in response to its biological environment and stimuli is becoming the foundation of many biomedical research fields, including drug development, nanotoxicity study, biomarker discovery, cancer diagnosis and treatment, and many other areas. However, there is no such instrument currently available to perform comprehensive, quantitative measurement of various key parameters inside a cell simultaneously. This project aims to develop a novel multi-functional single-cell analyzer that integrates the capability of in vitro measurements of a number of important intracellular parameters. The new cell analyzer will address a number of critical issues in biomedical research such as cell necrosis and apoptosis, stem cell differentiation, and cancer cell biology. Three specific aims are designed in this project, including: 1) Development of tapered-fiber microprobes and associated instrumentation for in situ measurement of intracellular pH, temperature, and Raman spectroscopy; 2) Integration of tapered-fiber microsensors with morphology and structure monitoring capabilities to construct the single-cell analyzer; and 3) Evaluation and demonstration of the multi-functional single-cell analyzer using cytotoxicity of nanomaterials as an application model. The proposed novel single-cell analyzer is uniquely enabled by a number of key innovations embedded in this project. Assembly-free micro- and nano-size sensor probes will be designed and fabricated on the distal end of optical fiber tapers that can be directly inserted into a livin cell for in situ, quantitative measurement of various important parameters with high resolution. The innovative integration of these microsensors with microscopic imaging tools will allow comprehensive, systematic studies of the cell responses to the bio-environment and stimuli. The use of cytotoxicity of nanoparticles as an example to evaluate and demonstrate the basic functions of the single-cell analyzer will also provide valuable information for further performance enhancement, system optimization, and expansion of applications. The proposed research will be conducted by a multidisciplinary team consisting of three professors at Missouri University of Science and Technology (Missouri S&T). All major equipment required for this project is available on Missouri S&T campus. The team members have complementing skills in biochemistry/bioanalysis, cytotoxicity of nanomaterials, optical micro/nano sensors and instrumentation, and analytical instrument design and implementation. The interdisciplinary project of life science, electrical engineering, and chemistry will foster strong collaborations among faculty and students in different fields, and strengthen the biomedical- related research and infrastructure at Missouri S&T.