微流控芯片-激光诱导荧光检测器的研制及核酸片段分离检测中应用

Development of Laser-induced Fluorescence Detector for Deoxyribonucleic Acid Fragments Seperation by Microfluidic Chip

采用自行设计的共焦式激光诱导荧光检测器(激发波长635nm,发射波长670nm),以cy5染料对检测器的空间分辨能力和灵敏度进行了测试,检出限达到10-9mol/L。建立了一种微流控芯片快速分离检测DNA片段的方法。以0.8%羟丙基甲基纤维素(HPMC)为筛分介质,以(噻唑橙单体)To-pro-3为荧光标记染料,在玻璃微流控芯片中实现了dsDNA片段的无胶筛分和激光诱导荧光检测,12条DNA片段在75s内得到分离。

A confocal laser-induced fluorescence detector, with Aez 635.0 nm and Aem 670.0 nm, was designed and constructed. The space resolution and sensitivity was examined. The detection limit of Cy5 dye （C37H42O10N3S2K） was 10^-9 mol/L. A method was developed for deoxyribonucleic acid （DNA） fragments electrophoresis separation on microfluidic chip. Fragments of dsDNA were separated with non-gel sieving electrophoresis on microfluidic chip, detected by laser-induced fluorescence detector using To-pro-3 dye （a derivative of cyanien dye thiazole orange, structure was shown in the paper）.