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用于 CE 的具有银镜涂层检测窗口的高灵敏度 LIF 检测器和共线系统的小角度光学偏折

A high-sensitivity LIF detector with silver mirror coating detection window and small-angle optical deflection from collinear system for CE.

机构信息

College of Chemistry, Sichuan University, Chengdu, P. R. China.

出版信息

Electrophoresis. 2013 Dec;34(22-23):3163-70. doi: 10.1002/elps.201300281. Epub 2013 Oct 31.

Abstract

An LIF detector was integrated into a CE system based on silver mirror coating detection window and small-angle optical deflection from collinear configuration. For this detection scheme, the incident light beam was focused on capillary through the edge of a lens, resulting in a small deflection angle that deviated 18° from the collinear configuration. Meanwhile, the excitation light and emitted fluorescence were effectively reflected by silver mirror coating at the detection window. The fluorescence was collected through the center of the same lens and delivered to a PMT in the vertical direction. In contrast to conventional collinear LIF detection systems, the fluorescence intensity was greatly enhanced and the background level was significantly eliminated. FITC and FITC-labeled amino acids were used as model analytes to evaluate the performance with respect to design factors of this system. The limit LOD was estimated to be 0.5 pM for FITC (S/N = 3), which is comparable to that of optimized confocal LIF systems. All the results indicate that the proposed detection scheme will be promising for development of sensitive and low-cost CE system.

摘要

将 LIF 检测器集成到基于银镜涂层检测窗口和共线小角度光偏折的 CE 系统中。对于这种检测方案,入射光束通过透镜的边缘聚焦到毛细管上,导致小的偏折角与共线配置相差 18°。同时,激发光和发射荧光通过检测窗口上的银镜涂层得到有效反射。荧光通过同一透镜的中心收集,并沿垂直方向传输到 PMT。与传统的共线 LIF 检测系统相比,荧光强度大大增强,背景水平显著消除。FITC 和 FITC 标记的氨基酸被用作模型分析物来评估该系统设计因素的性能。对于 FITC(S/N = 3),检测限估计为 0.5 pM,与优化的共聚焦 LIF 系统相当。所有结果表明,所提出的检测方案有望开发出灵敏且低成本的 CE 系统。

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