基于微流控芯片-ICP-MS技术在线研究单细胞的砷摄取及毒性

Arsenic has attracted worldwide attention as an environment contaminant and carcinogens Despite that there exist massive researches on arsenic toxicology by animal model and in vitro cultured cells, it still appears lack of studies at single cell level, which might provide more information due to the cell heterogeneity. The toxic effects of arsenic depend on the arsenic species, cellular uptake, and the intrinsic properties of cells. Analysis arsenic uptake and corresponding physical response at single cell level will represent cell to cell variations more precisely and improve our understanding on the toxic effects and mechanism of arsenic. This proposal aims at integrate microfluidic chip with inductively coupled plasma mass spectrometry (ICP-MS) to develop a single cell analyzing platform. The microfluidic chip will combine single cell sorting, manipulating, electrical measuring, optical monitoring functions to achieve on-line test of cell impedance and cell oxidative stress, furthermore the arsenic content of the cell would be examined in the ICP-MS; This proposal will analyze multiple influencing factors at single cell level including arsenic content, cell membrane state, cell oxidative stress to clarify cell heterogeneity related to arsenic cytotoxity. This Microfluidic-ICP-MS on-line test system will provide an alternative way to study arsenic exposure, uptake, metabolites and toxic mechanism

砷是一种广泛关注的污染物和致癌物，现有砷的毒理学研究数据主要来源于活体动物模型和离体细胞暴露实验。但砷的毒性效应取决于砷的形态、细胞对砷的摄入量及细胞的自身特性，从单细胞水平研究砷的摄取和影响因素以及胞内应激反应对深入理解砷的毒性效应和致毒机制具有重要意义。本项目将搭建微流控芯片与电感耦合等离子体质谱（ICP-MS）联用技术平台，利用双层阀控微流控芯片的单细胞分选能力及操作灵活性，结合细胞电性质测试方法和荧光检测技术，实现单细胞包裹、传输、砷暴露以及细胞膜间阻抗、细胞内活性氧水平的在线检测；并利用ICP-MS定量测定不同形态砷暴露后单细胞内砷的含量；综合细胞砷摄取量、细胞膜状态、细胞氧化应激等指标，在单细胞水平上阐明细胞异质性与砷摄取、砷细胞毒性的相关性。本项目所开发的微流控芯片-ICP-MS在线单细胞技术平台可为研究砷的暴露与摄取、体内代谢与形态转化、毒性效应与机制提供新的方法。