title: Efficient sample preparation from complex biological samples using a sliding lid for immobilized droplet extractions.
abstract: 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.
authors.affiliation: Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin , Madison, Wisconsin 53705, United States.
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Casavant
authors.forename: David J
authors.initials: DJ
authors.lastname: Guckenberger
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
authors.forename: Scott M
authors.initials: SM
authors.lastname: Berry
mesh_terms: D000818:Animals; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation*; D005591:Chemical Fractionation / Q000295:instrumentation*; D004247:DNA / Q000302:isolation & purification*; D004867:Equipment Design; D049452:Green Fluorescent Proteins / Q000302:isolation & purification; D006678:HIV / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008164:Luminescent Proteins / Q000302:isolation & purification; D059346:Magnets / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D000097573:Red Fluorescent Protein
chemical_list: D004247:DNA; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011506:Proteins; D012367:RNA, Viral; D000097573:Red Fluorescent Protein