时间分辨多色电化学发光余辉成像研究及多元分析应用

Multicolor electrochemiluminescence (ECL) imaging has become a very attractive research hotspot in the field of multiple luminescence analysis. However, it is difficult to achieve multicolor ECL imaging in the full wavelength only with the combination of ECL emitters. This research aims to develop ECL multi-sensor technology and improve the multicolor ECL theory system. A series of high-quality long persistence luminescent materials with tunable emission wavelength, size and afterglow time were introduced to achieve multicolor ECL imaging through converting single ECL signal into a superimposed signal. Moreover, The ECL signal was converted into a glow afterglow signal by using the principle of energy resonance transfer to achieve ECL afterglow imaging. The effects of emission wavelength, particle size, co-reactant reagent, spectral overlap and spatial distance on color superposition and resonance energy transfer were systematically investigated. The electroactive sites were indirectly reflected by the afterglow luminescent signal to avoid the interference of electroactive substances. A time coding strategy was proposed to develop a time-resolved multi-sensor platform for ECL imaging utilizing the differences of luminescent time for long persistence luminescent materials. This method could realize real-time and sensitive detection of complex tumor markers, providing a potent method for non-interfering and time-resolved ECL imaging analysis.

多色电致化学发光成像（ECLI）已成为多元发光分析领域十分引人注目的研究热点。但仅以ECL发光体组合较难实现全波段范围内的多色ECL成像。本项目以完善多色ECL理论体系，开发ECL多元传感技术为目标，在通过引入一系列发射波长、尺寸、余辉时间等可调的高质量长余辉发光材料的基础上，利用颜色叠加效应将单一ECL信号转变成叠加信号，实现多色ECL成像；利用共振能量转移原理，将ECL信号转变为余辉发光信号，实现ECL余辉成像。系统考察发射波长、粒子尺寸、共反应试剂、光谱重叠、空间距离对颜色叠加和能量共振转移的影响；以余辉发光信号间接反映的电活性位点，避开电活性物质干扰，从发光层面探讨ECL机理。利用长余辉材料持续发光时间不同，提出时间编码策略，开发时间分辨ECL成像多元传感平台，实现对复杂肿瘤标志物的实时、高灵敏检测，为无干扰、时间分辨ECL成像分析提供重要的方法学参考。

本项目以完善多色ECL理论体系，开发ECL多元传感技术为目标，在通过引入一系列发射波长、尺寸、余辉时间等可调的高质量长余辉发光材料的基础上，实现多色ECL和余辉ECL。系统考察发射波长、粒子尺寸、共反应试剂、光谱重叠、空间距离对颜色叠加和能量共振转移的影响。利用长余辉材料持续发光时间不同，提出时间编码策略，开发时间分辨ECL成像传感平台。主要从三个层面实现：（i）拓展除鲁米诺（蓝光）和联吡啶钌（橙光）之外的ECL试剂。我们开发了新型的半导体量子点ECL试剂和波长可调控的碳点ECL试剂。（ii）通过长余辉粒子的余辉颜色与常规ECL试剂叠加实现多色ECL。我们开发了一系列微纳米长余辉粒子，包括无机纳米粒子和长余辉碳点，可控合成了不同发射波长和余辉时间的 LAP 材料，并探索 LAP 材料的合成、组装新策略，制备余辉时间可调、能级可调、种类齐全的 LAP 材料，研究了其发光特性（发光形式、强度、激发光谱）与其结构、表面态及带隙能级的关系，完善了长余辉合成策略和理论体系，并成功开发出新型多色长余辉碳点，拓宽了LAP材料类别。探讨了长余辉纳米粒子ZGGO:Cr3+,Yb3+,Er3+和ECL试剂鲁米诺之间的相互作用，考察了发射波长、余辉强度、余辉时间、ECL 发光体种类的影响，构建了时间分辨分析体系，实现了肿瘤标志物传感检测应用。（iii）ECL试剂通过共振能量转移传递给长余辉材料，实现余辉ECL。我们直接以长余辉粒子作为ECL试剂，并通过控制ECL相关电化学过程中改变施加电压和脉冲时间实现余辉ECL。