单细胞中微量元素/形态与细胞相互作用及分析新方法研究

Single cell analysis is mostchallenging as amultidisciplinaryemerging research field including analytical chemistry, biology and medical science. The related chemical and biological information in cells is currently mainly achieved by the analysis of cell population due to the limitations of the detection techniques. However, the statistical average information from the cell population could hardly illustrate the differences between individual cells, and in many cases this will cause the loss of important information of biological and/or medical significance. At this junction, the present project will focus on the investigations on the interactions between trace chemical species and single cells as well as the development of novel detection protocols for their analysis. These will include: the development of novel sampling procedures for single cell, and the analysis of metal/metalloid in the related single cell; the exploitation of the regulation mechanism and metabolism behavior of the essential/non-essential elements (metal/metalloid), in order to further elucidate their conversion or variation of their chemical forms on the cell surface or in the cell interior; to investigate the interaction and labeling effects of new metal/metalloid nanoparticle probes with the cells of interest, and further to realize the rapid diagnoseof the variation of symptom for a specific disease (in the present study, we will focus on the Alzheimer's disease), due to the unbalanced (deficiency or excessive)essential elements for life; and finally to study the in-cell migration/transformation pathway and the metabolism behavior of metallic drugs (anti-cancer drugs and those for the treatment of Alzheimer's disease), for the purpose to elucidate the interaction mechanism of anti-cancer drugs and neoplasticcells.

单细胞分析是分析科学、生物学和医学渗透发展形成的跨学科前沿，是分析科学中极具挑战性的研究领域。目前，细胞中有关化学及生物学信息的获取多依赖细胞群体的分析。然而这种统计平均结果往往掩盖单个细胞间的差异，使具有重要生物学及医学意义的信息丢失。鉴于此，本项目拟开展单细胞中微量组份与细胞相互作用及其分析新方法的研究，包括：探索新型单细胞进样系统，建立单细胞中金属/类金属元素的检测方法；研究细胞/细菌中生命必需/非必需元素的生物平衡调控与代谢行为，探索其在细胞/细菌表面或胞内的转化及形态变化；探索新型金属/类金属纳米探针及其与细胞/细菌的作用及标记，用于快速诊断由于生命必需元素失衡（缺失或过量）而导致的相关疾病病征的变化（以阿尔茨海默氏病为研究对象）；研究金属类药物（抗癌药或阿尔茨海默氏病治疗药物）在细胞中的迁移转化与代谢行为，为从生命最基本单元（细胞）层面上揭示抗癌药物的作用机理提供依据。

单细胞分析是分析科学、生物学和医学渗透发展形成的跨学科前沿，是分析科学中极具挑战性的研究领域。目前，细胞中有关化学及生物学信息的获取多依赖细胞群体的分析。然而这种统计平均结果往往掩盖单个细胞间的差异，使具有重要生物学及医学意义的信息丢失。本项目探索性构建单细胞分离系统的可行性并构建基于商业化的四通接口、微流控芯片和毛细管等流体操控单元，构建了三种形式的单细胞分离系统。基于商业化四通接口，构建三维微交叉液滴发生系统对单细胞进行封装，封装有单细胞的液滴引入检测器进行检测。创新性的引入碳酸二甲酯作为连续相添加剂，可以满足后续无机质谱检测，不仅可以保证等离子体焰炬的稳定，而且可以极大的减少采样锥孔的碳沉积和有机质在检测过程中的负面作用；开发了Dean流诱导的细胞有序排列平台，用于从细胞悬浮液中聚焦单个细胞。悬浮细胞在螺旋毛细管中受流体惯性力和Dean力共同作用下，不断聚焦，形成单列细胞流。有序单细胞采样系统在保持高通量的同时，提供了更有利于单细胞检测的时空分辨率，同时避免了液滴封装在单细胞分析中遇到的有机相干扰问题；设计构建了具有螺旋通道的微流控芯片，同样在流体惯性力和Dean力共同作用下，实现单细胞的有效聚焦和分离，同时该结构还可以实现在较宽的流体速度范围内大浓度细胞悬液中的细胞自发有序地排列。研究了高通量单细胞中金属纳米粒子/元素的定量方法，金属类药物在（单）细胞中的迁移转化、代谢以及其与特定蛋白的相互结合与作用，探索了新型金属/类金属纳米探针对细胞或者特定生物分子的标记及定量检测方法，研究了（单）细胞中金属形态的分析定量方法。