Casavant Benjamin P, Guckenberger David J, Beebe David J, Berry Scott M
Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin , Madison, Wisconsin 53705, United States.
Anal Chem. 2014 Jul 1;86(13):6355-62. doi: 10.1021/ac500574t. Epub 2014 Jun 13.
Sample preparation is a major bottleneck in many biological processes. Paramagnetic particles (PMPs) are a ubiquitous method for isolating analytes of interest from biological samples and are used for their ability to thoroughly sample a solution and be easily collected with a magnet. There are three main methods by which PMPs are used for sample preparation: (1) removal of fluid from the analyte-bound PMPs, (2) removal of analyte-bound PMPs from the solution, and (3) removal of the substrate (with immobilized analyte-bound PMPs). In this paper, we explore the third and least studied method for PMP-based sample preparation using a platform termed Sliding Lid for Immobilized Droplet Extractions (SLIDE). SLIDE leverages principles of surface tension and patterned hydrophobicity to create a simple-to-operate platform for sample isolation (cells, DNA, RNA, protein) and preparation (cell staining) without the need for time-intensive wash steps, use of immiscible fluids, or precise pinning geometries. Compared to other standard isolation protocols using PMPs, SLIDE is able to perform rapid sample preparation with low (0.6%) carryover of contaminants from the original sample. The natural recirculation occurring within the pinned droplets of SLIDE make possible the performance of multistep cell staining protocols within the SLIDE by simply resting the lid over the various sample droplets. SLIDE demonstrates a simple easy to use platform for sample preparation on a range of complex biological samples.
样品制备是许多生物过程中的一个主要瓶颈。顺磁性颗粒（PMPs）是从生物样品中分离目标分析物的一种普遍方法，因其能够充分取样溶液并易于用磁铁收集而被使用。PMPs用于样品制备主要有三种方法：（1）从与分析物结合的PMPs中去除液体；（2）从溶液中去除与分析物结合的PMPs；（3）去除底物（连同固定有与分析物结合的PMPs）。在本文中，我们使用一种称为固定液滴萃取滑动盖（SLIDE）的平台，探索基于PMPs的样品制备的第三种且研究最少的方法。SLIDE利用表面张力和图案化疏水性原理，创建了一个易于操作的平台，用于样品分离（细胞、DNA、RNA、蛋白质）和制备（细胞染色），无需耗时的洗涤步骤、使用不混溶的液体或精确的固定几何形状。与使用PMPs的其他标准分离方案相比，SLIDE能够以低（0.6%）的原始样品污染物残留率进行快速样品制备。SLIDE固定液滴内自然发生的再循环使得通过简单地将盖子放置在各种样品液滴上，就能在SLIDE内执行多步细胞染色方案。SLIDE展示了一个用于一系列复杂生物样品制备的简单易用平台。
