Microfluidic active sorting of DNA molecules labeled with single quantum dots using flow switching by a hydrogel sol-gel transition
Microfluidic active sorting of DNA molecules labeled with single quantum dots using flow switching by a hydrogel sol-gel transition
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DOI：
10.1016/j.snb.2011.06.043
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发表时间：
2011-11-28
影响因子：
8.4
通讯作者：
Funatsu, Takashi
中科院分区：
文献类型：
--
作者：
Haneoka, Mai;Shirasaki, Yoshitaka;Funatsu, Takashi
荧光激活细胞分选仪为生命科学做出了重大贡献。然而，该仪器具有局限性，包括无法检测并因此分选纳米尺寸的颗粒，例如量子点（Qdots）和病毒颗粒。在这里，已经开发了用于分析和分选纳米尺寸颗粒的微流体装置。为了实现灵敏的检测，样品流被流体动力学地护套，并有效地激发与聚焦激光束。通过热可逆凝胶化聚合物的溶胶-凝胶转变进行流动控制。这种流动控制方法使我们能够限制纳米级颗粒的横向扩散，并在类似于10 μ m的通道中对颗粒进行分选。在约4 mm/s的线流速下检测到直径接近10 nm的单个量子点，并且量子点被分选系统成功分选。在应用中使用开发的分选器，Qdot标记的肌动蛋白DNA与不需要的甘油醛-3-磷酸脱氢酶DNA分离，并且Qdot标记的肌动蛋白DNA的纯度在分选后增加。这项研究代表了用单个10 nm大小的量子点颗粒标记的生物分子的主动分离的第一个例子。(C)2011 Elsevier B. V.保留所有权利。
The fluorescence-activated cell sorter instrument has contributed significantly to life sciences. However, this instrument has limitations including the inability to detect and hence sort nanometer-sized particles such as quantum dots (Qdots) and virus particles. Here, a microfluidic device for analyzing and sorting nanometer-sized particles has been developed. To achieve sensitive detection, the sample flow was hydrodynamically sheathed and effectively excited with a focused laser beam. Flow control was performed by a sot-gel transition of a thermoreversible gelation polymer. This flow control approach enabled us to restrict the lateral diffusion of nanometer-sized particles and to sort the particles in similar to 10 mu m channels. Single Qdots with diameters of similar to 10 nm were detected at a linear flow velocity of about 4 mm/s, and the Qdots were successfully sorted with the sorting system. Using the developed sorter in an application, Qdot-labeled actin DNA was separated from unwanted glyceraldehyde-3-phosphate dehydrogenase DNA, and the purity of Qdot-labeled actin DNA increased following the sorting. This study represents the first example of active separation of biomolecules labeled with single 10 nm-sized particles of Qdots. (C) 2011 Elsevier B.V. All rights reserved.