Two-dimensional direct-reading fluorescence spectrograph for DNA sequencing by capillary array electrophoresis
Two-dimensional direct-reading fluorescence spectrograph for DNA sequencing by capillary array electrophoresis
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DOI：
10.1021/ac001001c
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发表时间：
2001-03-15
影响因子：
7.4
通讯作者：
Dovichi, NJ
中科院分区：
文献类型：
--
作者：
Zhang, JZ;Yang, MJ;Dovichi, NJ
我们报告了一种紧凑的二维直读荧光光谱仪，并展示了其在毛细管电泳 DNA 测序中的应用。该检测比色皿基于鞘流，其中毛细管以二维阵列终止于充满液体的室中，该室用缓冲液加压。薄金属板位于毛细管下游。该挡板具有一系列孔，与毛细管的位置精确匹配。缓冲液流过这些孔，从毛细血管中吸取分析物，形成清晰的细丝阵列。椭圆形激光束在上室中激发荧光。底部室用玻璃窗密封并从芯片侧排水。通过对穿过挡板孔的照明样品流进行成像来检测荧光。棱镜用于将每个样品的荧光分散到 CCD 相机上，以便同时监测每个毛细管的发射光谱。该仪器以 32 根毛细管配置进行演示，但可以扩展到数千根毛细管。
We report a compact, two-dimensional direct-reading fluorescence spectrograph and demonstrate its application to DNA sequencing by capillary away electrophoresis. The detection cuvette is based on sheath flow, wherein the capillaries terminate in a two-dimensional array in a fluid-filled chamber that is pressurized with buffer. A thin metal plate is located downstream from the capillaries. This barrier plate has an array of holes that precisely matches the location of the capillaries. Buffer flows through the holes, drawing analyte from the capillaries in a well-defined array of thin filaments. Fluorescence is excited in the upper chamber with an elliptically shaped laser beam. The bottom chamber is sealed with a glass window and drained from die side. Fluorescence is detected by imaging the illuminated sample streams through the holes in the barrier plate. A prism is used to disperse fluorescence from each sample across a CCD camera so that the emission, spectrum is monitored simultaneously from each capillary. The instrument is demonstrated in a 32-capillary configuration but can be scaled to several thousand capillaries.