Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, V5A 1S6 BC, Canada.
Anal Chim Acta. 2011 Feb 14;687(1):12-27. doi: 10.1016/j.aca.2010.11.056. Epub 2010 Dec 17.
Microarray DNA hybridization techniques have been used widely from basic to applied molecular biology research. Generally, in a DNA microarray, different probe DNA molecules are immobilized on a solid support in groups and form an array of microspots. Then, hybridization to the microarray can be performed by applying sample DNA solutions in either the bulk or the microfluidic manner. Because the immobilized probe DNA binds and retains its complementary target DNA, detection is achieved through the read-out of the tagged markers on the sample target molecules. The recent microfluidic hybridization method shows the advantages of less sample usage and reduced incubation time. Here, sample solutions are confined in microfabricated channels and flow through the probe microarray area. The high surface-to-volume ratio in microchannels of nanolitre volume greatly enhanced the sensitivity as obtained with the bulk solution method. To generate nanolitre flows, different techniques have been developed, and this including electrokinetic control, vacuum suction and syringe pumping. The latter two are pressure-driven methods which are more flexible without the need of considering the physicochemical properties of solutions. Recently, centrifugal force is employed to drive liquid movement in microchannels. This method utilizes the body force from the liquid itself and there are no additional solution interface contacts such as from electrodes or syringes and tubing. Centrifugal force driven flow also features the ease of parallel hybridizations. In this review, we will summarize the recent advances in microfluidic microarray hybridization and compare the applications of various flow methods.
微阵列 DNA 杂交技术已广泛应用于基础和应用分子生物学研究。通常,在 DNA 微阵列中,不同的探针 DNA 分子被固定在固体载体上并形成微点阵列。然后,可以通过以 bulk 或微流控的方式应用样品 DNA 溶液进行杂交。由于固定的探针 DNA 与互补的靶 DNA 结合并保留,因此通过读取样品靶分子上标记的标记物来实现检测。最近的微流控杂交方法显示出减少样品使用量和缩短孵育时间的优点。在这里,样品溶液被限制在微加工通道中并流过探针微阵列区域。纳升体积的微通道中高的表面积与体积比极大地提高了与 bulk 溶液方法相比的灵敏度。为了产生纳升流量,已经开发了不同的技术,包括电动控制、真空抽吸和注射器泵。后两种是压力驱动的方法,不需要考虑溶液的物理化学性质,因此更加灵活。最近,离心力被用于驱动微通道中的液体运动。这种方法利用液体本身的体力,没有额外的溶液界面接触,如电极或注射器和管道。离心力驱动的流动还具有易于进行并行杂交的特点。在这篇综述中,我们将总结微流控微阵列杂交的最新进展,并比较各种流动方法的应用。