纳米孔道技术在环境污染物单细胞毒性研究中的应用

Recently, the biological toxicity of environmental pollutants has caused great threat to the ecological environment and public health. The toxicity analysis of pollutants, especially cytotoxicity analysis, has become an important topic in the fields of environmental chemistry and biomedicine science. Traditional cell analysis methods such as cell lysis would cause irreversible damage to cell function, which could hardly acquire the dynamic changes of intracellular molecules. Cytotoxicity analysis of pollutants at single living cells could help to better understand the complex processes of cell damage/death and reveal cellular heterogeneity. This project aims to develop a glass nanopore based method which could direct inject pollutants into single cells with subcellular resolution. Moreover, very small volumes of cellular content could be extracted without causing damage to the living cells, which could be analyzed by Time-of-Flight Secondary Ion Mass Spectrometry. This single cell analysis method will be ultimately applied in evaluating the cellular toxicity of pollutants.

近年来，环境污染已经成为威胁全球生态环境和公众健康的头号杀手之一，污染物的生物毒性分析，尤其是细胞毒性分析已成为环境化学及生物医学等领域的研究热点。传统细胞分析方法如细胞裂解等手段会对细胞功能造成不可逆损伤，无法获得单细胞内生物活性分子的动态变化。在单细胞水平上研究环境污染物的生物毒性有助于揭示细胞异质性，理解细胞损伤或死亡的动态复杂过程。本项目针对环境污染物单细胞毒性这一研究难题，拟通过玻璃纳米孔道的可控针尖实现污染物在亚细胞水平上的定向灌注；进一步构建纳米孔道与飞行时间-二次离子质谱相结合的单细胞分析系统，实现单细胞内微量液体组分的提取分析，实时监测细胞内多种组分的动态变化，以期在单细胞水平上实现环境污染物毒性研究，并对其毒性机理进行分析。

本项目针对纳米孔道技术在环境污染物单细胞毒性研究中的应用，设计制备了可用于单细胞分析的玻璃纳米孔道传感界面，发展了新型孔道电极界面用于解析环境污染物AgNPs在单颗粒水平上的氧化过程，为其细胞毒性解析奠定基础；发展了基于飞行时间-二次离子质谱的单细胞组学分析方法，实现了疾病标志物的有效识别；进一步利用玻璃纳米孔道实现超微量液体的精准操控，可用于单个活细胞的外源物质定向灌注以及细胞质的有效提取，实现单个活细胞的飞行时间-二次离子质谱研究，从而为单细胞组学技术的发展以及环境污染物单细胞毒性研究提供新方法。