高通量CE系统的多波长荧光检测

Shih-wei Lin, G. Chang, Che-Hsin Lin
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引用次数: 2

摘要

本文提出了一种用于生物样品高通量分析的多波长荧光检测新方法。一般来说,荧光染料可以用特定波长的光源激发,然后被激发的荧光发出波长较长的光,因此需要在荧光显微镜上设置特殊的滤光片。本研究没有采用传统的激光诱导荧光方案,而是采用双聚光照明光源进行荧光激发,采用UV-VIS-NIR光谱仪进行发射信号检测。用这种方法可以在单个CE通道中同时激发和检测多个荧光染料。此外,由于不需要复杂的光学滤光片和激光源来进行检测,因此所提出的系统简单且成本低。实验结果表明,该系统对混合荧光样品的单次并行检测是可行的。荧光染料包括Atto 610、Rhodamine B和FITC,成功地同时检测和鉴定。该系统检测FITC荧光的LOD(检测限)可低至10-5 M(信噪比=5.56),适用于一般生物分析应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Multi-Wavelength Fluorescence Detection for a High-Throughput CE System under a Spectrum Equipped Diascopic Microscope Configuration
This paper presents a novel method for multi-wavelength fluorescence detection for high-throughput analysis of bio-samples in a micro-CE chip. In general, fluorescent dye can be excited with light sources in a specific wavelengths then the excited fluorescence emits light with longer wavelengths such that a special filter set on a fluorescence microscope is required for this application. Instead of using conventional laser induced fluorescence scheme, this study adopts a diascopic illumination light source for fluorescence excitation and an UV-VIS-NIR spectrometer for emission signals detection. Multiple fluorescence dyes can be excited and detected simultaneously in a single CE channel with this approach. In addition, the proposed system is simple and low-cost since no sophisticated optical filter sets and laser sources are required for the detection purpose. Experimental result shows that the proposed system is feasible for parallel detecting a mixed fluorescence sample in a single run. Fluorescence dyes including Atto 610, Rhodamine B and FITC are successfully detected and identified simultaneously. The LOD (limit of detection) for detecting FITC fluorescence of the proposed system can be as low as 10-5 M(SNR=5.56) which is applicable for to general bio-analytical applications.
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