Li Hai-Fang, Lin Jin-Ming, Su Rong-Guo, Uchiyama Katsumi, Hobo Toshiyuki
Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, China.
Electrophoresis. 2004 Jun;25(12):1907-15. doi: 10.1002/elps.200305867.
A simple and easy-to-use integrated laser-induced fluorescence detector for microchip electrophoresis was constructed and evaluated. The fluid channels and optical fiber channels in the glass microchip were fabricated using standard photolithographic techniques and wet chemical etching. A 473 nm diode-pumped laser was used as the excitation source, and the collimation and collection optics and mirrors were discarded by using a multimode optical fiber to couple the excitation light straight into the microchannel and placing the microchip directly on the top of the photomultiplier tube. A combination of filter systems was incorporated into a poly(dimethylsiloxane) layer, which was reversibly sealed to the bottom of the microchip to eliminate the scattering excitation light reaching to the photomultiplier tube. Fluorescein/calcein samples were taken as model analytes to evaluate the performance with respect to design factors. The detection limits were 0.05 microM for fluorescein and 0.18 microM for calcein, respectively. The suitability of this simple detector for fluorescence detection was demonstrated by baseline separation of fluorescein isothiocyanate (FITC)-labeled arginine, phenylalanine, and glycine and FITC within 30 s at separation length of 3.8 cm and electrical field strength of 600 V/cm.
构建并评估了一种用于微芯片电泳的简单易用的集成激光诱导荧光检测器。玻璃微芯片中的流体通道和光纤通道采用标准光刻技术和湿化学蚀刻工艺制造。使用473 nm二极管泵浦激光器作为激发源，通过使用多模光纤将激发光直接耦合到微通道中，并将微芯片直接放置在光电倍增管顶部，从而省去了准直和收集光学器件及镜子。将一组滤光系统集成到聚二甲基硅氧烷层中，该层可逆地密封在微芯片底部，以消除到达光电倍增管的散射激发光。以荧光素/钙黄绿素样品作为模型分析物，评估其在设计因素方面的性能。荧光素和钙黄绿素的检测限分别为0.05 μM和0.18 μM。在3.8 cm的分离长度和600 V/cm的电场强度下，异硫氰酸荧光素（FITC）标记的精氨酸、苯丙氨酸、甘氨酸和FITC在30 s内实现基线分离，证明了这种简单检测器适用于荧光检测。
