生物体系自助式原位荧光信号放大平台的创建及生化应用

In situ, real-time acquisition of chemical and biological information is the cornerstone of exploring the essence of biological process, and is also one of the core content of researches in analytical chemistry. This project is based on in situ biochemical information acquisition at cellular and in vivo levels, trying to deal with the challenges of in situ detection of low abundance bioactive substances and major disease markers, focusing on breaking through the limitation of conventional fluorescent probe design and signal amplification methods to launch the fundamental research with original innovation：It is proposed for the first time to employ intracellular high abundance components as amplification units instead of exogenous substances, to realize the development of self-assistant in situ signal amplification strategy in biological system, thus creating ultra-sensitive visual analysis system for in situ fluorescence detection at cellular and in vivo levels, which hold the potential to be applied in the research of cell signal transduction, cell apoptosis, pancreatic beta cell injury and other physiological or pathological phenomenon, as well as revealing the molecular mechanisms behind pathologic process in major disease. The execution of the project will provide new ideas for the construction and development of high-efficiency in vivo fluorescence imaging probes, and give new tool and new method on molecular level and nano scale to explore the relation between the variation of biological active substance and mechanisms underlying life activities, as well as the connection between the level of key markers and causes of major diseases at cellular and in vivo levels.

原位、实时获取生化信息是探究生命现象本质的基石，也是当前分析化学研究核心内容之一。本项目以活细胞和活体内生化信息原位、实时获取为研究目标，应对生物体内低丰度生命活性物种和重大疾病关键标志物难于原位、实时检测的挑战，突破传统荧光探针设计思路和已有信号放大的局限性，开展原始创新研究：首次提出利用细胞内高丰度固有组分替代外源性物质作为信号放大单元，基于荧光染料和细胞高丰度组分的相互作用/反应，提出生物体系自助式原位信号放大新概念，创建细胞活体原位超灵敏荧光成像平台，并应用于细胞信号传导、细胞凋亡、损伤等生理或病理现象的分子作用研究，以此揭示隐藏在重大疾病病变过程背后的分子调控机制。本项目的执行将为丰富与发展高效活体荧光成像提供新思路，为探究细胞活体层面和纳米尺度与分子水平上生物活性物质变化与生命活动规律或重大疾病关键标志物与疾病成因之间关系提供新工具和新方法。

本项目以活细胞和活体内生化信息获取为出发点，应对生物体内低丰度生命活性物种和重大疾病关键标志物难于原位、实时检测的挑战，按照预定研究计划，开展生物体系自助式原位荧光放大新方法研究。研究团队经过五年同心协力，筛选优化了适用于高保真生物成像应用的细胞组分增强荧光染料分子，发展了高特异性分子识别单元和传感界面，创建了通用且生物相容的高灵敏活体荧光传感平台，获得多个生命活性物种和疾病关键标志物荧光放大成像探针，系列研究工作丰富了活体分析方法学，拓展了荧光信号放大策略应用范围，为从细胞和活体层面上发现和阐明痕量生物活性物质或疾病标志物变化规律与生命活动与疾病成因之间关系提供了新工具。项目执行期内，相关研究结果发表标注基金号SCI论文67篇（包括J. Am. Chem. Soc.2篇, Angew. Chem. Int. Ed.7篇, CCS. Chem.2篇, Chem. Sci.5篇, Anal. Chem.14篇），公开国家发明专利19项（授权15项），获湖南省自然科学一等奖1项。项目执行期间，项目组成员赴美国、加拿大、和新加坡等海外高校访学，组织承办了中国化学会第二十一届全国有机分析及生物分析学术研讨会，项目负责人受邀国内外学术会议报告14次，培养了青年教师8人，其中1人获国家优秀青年基金资助、1人获国家高层次人才计划青年项目资助、3人入选湖南省高层次人才计划、3人晋升为正教授、5人晋升为副教授，博士后3人，毕业研究生54人（博士10人，硕士44人）。