Microfluidic reactors for advancing the MS analysis of fast biological responses

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dc.contributor.authorLazar, Iuliana M.en
dc.contributor.authorDeng, J.en
dc.contributor.authorStremler, Mark A.en
dc.contributor.authorAhuja, Shreyaen
dc.contributor.departmentMechanical Engineeringen
dc.contributor.departmentBiological Sciencesen
dc.contributor.departmentFralin Biomedical Research Instituteen
dc.date.accessioned2019-03-29T13:05:27Zen
dc.date.available2019-03-29T13:05:27Zen
dc.date.issued2019-02-11en
dc.description.abstractThe response of cells to physical or chemical stimuli is complex, unfolding on time-scales from seconds to days, with or without de novo protein synthesis, and involving signaling processes that are transient or sustained. By combining the technology of microfluidics that supports fast and precise execution of a variety of cell handling operations, with that of mass spectrometry detection that facilitates an accurate and complex characterization of the protein complement of cells, in this work, we developed a platform that supports (near) real-time sampling and proteome-level capturing of cellular responses to a perturbation such as treatment with mitogens. The geometric design of the chip supports three critical features: (a) capture of a sufficient number of cells to meet the detection limit requirements of mass spectrometry instrumentation, (b) fluid delivery for uniform stimulation of the resident cells, and (c) fast cell recovery, lysis and processing for accurate sampling of time-sensitive cellular responses to a stimulus. COMSOL simulations and microscopy were used to predict and evaluate the flow behavior inside the microfluidic device. Proteomic analysis of the cellular extracts generated by the chip experiments revealed that the identified proteins were representative of all cellular locations, exosomes, and major biological processes related to proliferation and signaling, demonstrating that the device holds promising potential for integration into complex lab-on-chip work-flows that address systems biology questions. The applicability of the chips to study time-sensitive cellular responses is discussed in terms of technological challenges and biological relevance. © 2019, The Author(s).en
dc.description.notesThis work was supported by the National Science Foundation and in part by the National Institute of General Medical Sciences, through awards NSF/DBI-1255991 and NIGMS-1R01GM121920-01A1 to IML. We thank the OSU Mass Spectrometry Center for analyzing the proteomic samples with the Orbitrap Fusion Lumos mass spectrometer (a purchase made possible by instrumentation grant S10 OD020111 to C. Maier).en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1038/s41378-019-0048-3en
dc.identifier.issn2055-7434en
dc.identifier.issue1en
dc.identifier.other7en
dc.identifier.urihttp://hdl.handle.net/10919/88746en
dc.identifier.volume5en
dc.language.isoen_USen
dc.publisherNature Publishing Groupen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.titleMicrofluidic reactors for advancing the MS analysis of fast biological responsesen
dc.title.serialMicrosystems and Nanoengineeringen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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