Jiang G, Attiya S, Ocvirk G, Lee W E, Harrison D J
Department of Chemistry, University of Alberta, Edmonton, Canada.
Biosens Bioelectron. 2000 Jan;14(10-11):861-9. doi: 10.1016/s0956-5663(99)00056-1.
A highly sensitive laser induced fluorescence (LIF) detection system based on a 635 nm laser diode and cyanine-5 (Cy-5) dye, is described for use with a planar, microfluidic, capillary electrophoresis (CE) chip. The CE-chip is able to determine a protein biological threat agent simulant, ovalbumin (Ov), by performing an immunoassay separation of Cy-5 labeled anti-ovalbumin from its complex with Ov, in under 30 s. A confocal, epiluminescent detection system utilizing a photomultiplier tube gave optimum results with a 400 microm pinhole, an Omega 682DF22 emission filter, a 645DRLP02 dichroic mirror, a 634.54 +/- 5 nm excitation filter, and a Power Technology ACMO8 635 nm laser operated at 11.2 mW. Using this detector, a microchip CE device with a separation efficiency of 42,000 plates and an etch depth of 20 microm, gave a concentration detection limit of 9 pM Cy-5. This limit corresponds to the determination of 4560 injected molecules and detection of 900 of these molecules, given a probe volume of 1.6 pl and a probing efficiency of 20%.
描述了一种基于635 nm激光二极管和花菁-5（Cy-5）染料的高灵敏度激光诱导荧光（LIF）检测系统，用于与平面微流控毛细管电泳（CE）芯片配合使用。该CE芯片能够通过在30秒内对Cy-5标记的抗卵清蛋白与其与卵清蛋白（Ov）的复合物进行免疫分析分离，来测定蛋白质生物威胁模拟物卵清蛋白（Ov）。利用光电倍增管的共焦落射荧光检测系统，采用400微米针孔、Omega 682DF22发射滤光片、645DRLP02二向色镜、634.54±5 nm激发滤光片以及功率为11.2 mW的Power Technology ACMO8 635 nm激光器时，可获得最佳结果。使用该检测器，分离效率为42,000塔板且蚀刻深度为20微米的微芯片CE装置，Cy-5的浓度检测限为9 pM。给定探针体积为1.6皮升且探测效率为20%，此检测限对应于4560个注入分子的测定以及其中900个分子的检测。
