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Monitoring cell secretions on microfluidic chips using solid-phase extraction with mass spectrometry

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Abstract

Microfluidics is an enabling technology for both cell biology and chemical analysis. We combine these attributes with a microfluidic device for on-line solid-phase extraction (SPE) and mass spectrometry (MS) analysis of secreted metabolites from living cells in culture on the chip. The device was constructed with polydimethylsiloxane (PDMS) and contains a reversibly sealed chamber for perfusing cells. A multilayer design allowed a series of valves to control an on-chip 7.5 μL injection loop downstream of the cell chamber with operation similar to a six-port valve. The valve collects sample and then diverts it to a packed SPE bed that was connected in-line to treat samples prior to MS analysis. The valve allows samples to be collected and injected onto the SPE bed while preventing exposure of cells to added back pressure from the SPE bed and organic solvents needed to elute collected chemicals. Here, cultured murine 3T3-L1 adipocytes were loaded into the cell chamber and non-esterified fatty acids (NEFAs) that were secreted by the cells were monitored by SPE-MS at 30 min intervals. The limit of detection for a palmitoleic acid standard was 1.4 μM. Due to the multiplexed detection capabilities of MS, a variety of NEFAs were detected. Upon stimulation with isoproterenol and forskolin, secretion of select NEFAs was elevated an average of 1.5-fold compared to basal levels. Despite the 30-min delay between sample injections, this device is a step towards a miniaturized system that allows automated monitoring and identification of a variety of molecules in the extracellular environment.

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Acknowledgments

This research was funded by NIH R37 DK046960 to RTK. JPG was supported by NIH F32 EB019800. Spray tips and PicoClear unions were generously provided by New Objective Inc. TOF-MS experiments utilized Core Services of the Michigan Regional Comprehensive Metabolomics Resource Core (MRC2) supported by grant DK097153 of NIH to the University of Michigan.

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Authors and Affiliations

  1. Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA

    Colleen E. Dugan, James P. Grinias & Robert T. Kennedy

  2. Department of Molecular & Integrative Physiology, University of Michigan, Ann Arbor, MI, 48109, USA

    Sebastian D. Parlee

  3. Department of Internal Medicine, University of Michigan, Ann Arbor, MI, 48109, USA

    Mahmoud El-Azzouny & Charles R. Evans

  4. Department of Pharmacology, University of Michigan, Ann Arbor, MI, 48109, USA

    Robert T. Kennedy

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  1. Colleen E. Dugan
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  2. James P. Grinias
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Correspondence to Robert T. Kennedy.

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Dugan, C.E., Grinias, J.P., Parlee, S.D. et al. Monitoring cell secretions on microfluidic chips using solid-phase extraction with mass spectrometry. Anal Bioanal Chem 409, 169–178 (2017). https://doi.org/10.1007/s00216-016-9983-0

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