迈向聚二甲基硅氧烷（PDMS）微器件中的单分子分析：从超低染料浓度检测到单DNA分子研究
Towards single molecule analysis in PDMS microdevices: from the detection of ultra low dye concentrations to single DNA molecule studies.
作者信息
Ros A, Hellmich W, Duong T, Anselmetti D
机构信息
Experimental Biophysics, Physics Faculty, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany.
出版信息
J Biotechnol. 2004 Aug 26;112(1-2):65-72. doi: 10.1016/j.jbiotec.2004.04.020.
In this paper, we report on the performance of electrophoretical separation and laser-induced fluorescence (LIF) detection of dyes and fluorescently labeled biomolecules in poly(dimethylsiloxane) (PDMS) microdevices. The dyes fluorescein and fluorescein isothiocyanate (FITC) have been separated effectively in nM concentrations. Fluorescein injections gave linear concentration response in the range from 4 to 100 pM. As ultimate detection sensitivity, 100 fM injected fluorescein was obtained. Further, 100 fM injected fluorescein could be detected. This is to our knowledge the smallest electrokinetically injected dye concentration detected on a microchip. Injection studies of fluorescently labeled avidin revealed a theoretical detection limit of 25 nM for laser-induced fluorescence detection in good agreement with separations in glass chips. Furthermore, the injection of several and even one single DNA molecule using a PDMS cross injector has been demonstrated as well as free solution separation of lambda- and T2-DNA (60 pM each) in periodically structured channels.
在本文中，我们报告了在聚二甲基硅氧烷（PDMS）微器件中对染料和荧光标记生物分子进行电泳分离及激光诱导荧光（LIF）检测的性能。荧光素和异硫氰酸荧光素（FITC）两种染料在纳摩尔浓度下已得到有效分离。荧光素进样在4至100皮摩尔范围内给出线性浓度响应。作为最终检测灵敏度，获得了100飞摩尔进样荧光素。此外，100飞摩尔进样荧光素能够被检测到。据我们所知，这是在微芯片上检测到的最小的电动进样染料浓度。对荧光标记抗生物素蛋白的进样研究表明，激光诱导荧光检测的理论检测限为25纳摩尔，这与在玻璃芯片上的分离结果吻合良好。此外，还展示了使用PDMS交叉进样器进样几个甚至单个DNA分子，以及在周期性结构通道中对λ-DNA和T2-DNA（各60皮摩尔）进行自由溶液分离。
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