Institute of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan.
Biosens Bioelectron. 2010 Oct 15;26(2):717-22. doi: 10.1016/j.bios.2010.06.056. Epub 2010 Jul 3.
A simple method is proposed for modulating the excitation light used for multi-color fluorescence detection in a single capillary electrophoresis (CE) channel. In the proposed approach, a low-cost commercial liquid crystal device (LCD) projector with digitally-modulated LCD switches is used to provide the illumination light source and the fluorescence emitted from the CE chip is synchronously detected using an ultraviolet-visible-near infrared (UV-vis-NIR) spectrometer. The modulated light source enables the detection of multiple fluorescence signals within a single CE channel without the need of mechanically switching optical components. In order to enhance the sensing performance of the proposed system, two short-pass filters and one band-pass filter are inserted into the LCD projector to modify the wavelength spectra for fluorescence excitation. With this simple approach, the signal-to-noise (SN) ratio of the fluorescence detection signals is greatly improved by a factor of approximately 22 when detecting Atto647N fluorescent dye. The feasibility of the proposed multi-color CE detection approach is demonstrated by detecting two different samples including a mixed sample comprising FITC, Rhodamine B and Atto647N fluorescent dyes and a bio-sample composed of two ssDNAs labeled with FITC and Cy3, respectively. Results confirm that the digitally-modulated excitation system proposed in this study has significant potential for the parallel analysis of fluorescently-labeled bio-samples using a multi-color detection scheme.
提出了一种用于调制单根毛细管电泳 (CE) 通道中多色荧光检测用激发光的简单方法。在提出的方法中,使用低成本的商用液晶器件 (LCD) 投影仪和数字调制的 LCD 开关提供照明光源,并用紫外-可见-近红外 (UV-vis-NIR) 分光光度计同步检测从 CE 芯片发出的荧光。调制光源使得能够在单个 CE 通道内检测多个荧光信号,而无需机械切换光学组件。为了增强所提出系统的传感性能,在 LCD 投影仪中插入两个短通滤波器和一个带通滤波器,以修改荧光激发的波长谱。通过这种简单的方法,在检测 Atto647N 荧光染料时,荧光检测信号的信噪比 (SN) 大大提高了约 22 倍。通过检测包括 FITC、Rhodamine B 和 Atto647N 荧光染料的混合样品以及分别用 FITC 和 Cy3 标记的两种 ssDNA 的生物样品,证明了所提出的多色 CE 检测方法的可行性。结果证实,本研究中提出的数字调制激发系统具有使用多色检测方案对荧光标记生物样品进行并行分析的巨大潜力。