title: High-throughput microfluidic device for single cell analysis using multiple integrated soft lithographic pumps.
abstract: The ability to accurately control fluid transport in microfluidic devices is key for developing high-throughput methods for single cell analysis. Making small, reproducible changes to flow rates, however, to optimize lysis and injection using pumps external to the microfluidic device are challenging and time-consuming. To improve the throughput and increase the number of cells analyzed, we have integrated previously reported micropumps into a microfluidic device that can increase the cell analysis rate to ∼1000 cells/h and operate for over an hour continuously. In order to increase the flow rates sufficiently to handle cells at a higher throughput, three sets of pumps were multiplexed. These pumps are simple, low-cost, durable, easy to fabricate, and biocompatible. They provide precise control of the flow rate up to 9.2 nL/s. These devices were used to automatically transport, lyse, and electrophoretically separate T-Lymphocyte cells loaded with Oregon green and 6-carboxyfluorescein. Peak overlap statistics predicted the number of fully resolved single-cell electropherograms seen. In addition, there was no change in the average fluorescent dye peak areas indicating that the cells remained intact and the dyes did not leak out of the cells over the 1 h analysis time. The cell lysate peak area distribution followed that expected of an asynchronous steady-state population of immortalized cells.
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS, USA.
authors.forename: Damith E W
authors.initials: DE
authors.lastname: Patabadige
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS, USA.
authors.forename: Tom
authors.initials: T
authors.lastname: Mickleburgh
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS, USA.
authors.forename: Lorin
authors.initials: L
authors.lastname: Ferris
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS, USA.
authors.forename: Gage
authors.initials: G
authors.lastname: Brummer
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS, USA.
authors.forename: Anne H
authors.initials: AH
authors.lastname: Culbertson
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS, USA.
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Culbertson
mesh_terms: D002264:Carboxylic Acids; D002469:Cell Separation; D004586:Electrophoresis; D005452:Fluoresceins; D005456:Fluorescent Dyes; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D059010:Single-Cell Analysis; D013601:T-Lymphocytes
chemical_list: D002264:Carboxylic Acids; D005452:Fluoresceins; D005456:Fluorescent Dyes; C436864:Oregon Green 488 carboxylic acid; C027780:carboxyfluoresceindiacetate
keywords: Electrophoresis; Multilayer microfluidic devices; Peristaltic pumps; Single cell analysis