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A well-mixed, polymer-based microbioreactor with integrated optical measurements

Authors

  • Zhiyu Zhang,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Mass. Ave., 66-566, Cambridge, Massachusetts 02139; telephone: +1(617)253-4589; fax: +1(617)258-8224
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  • Nicolas Szita,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Mass. Ave., 66-566, Cambridge, Massachusetts 02139; telephone: +1(617)253-4589; fax: +1(617)258-8224
    Current affiliation:
    1. MIC, Department of Micro and Nanotechnology, Technical University of Denmark (DTU), DK-2800 Kgs. Lyngby, Denmark.
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  • Paolo Boccazzi,

    1. Department of Biology and Health Sciences and Technology, Massachusetts Institute of Technology, 77 Mass. Ave., 68-370A, Cambridge, Massachusetts 02139
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  • Anthony J. Sinskey,

    1. Department of Biology and Health Sciences and Technology, Massachusetts Institute of Technology, 77 Mass. Ave., 68-370A, Cambridge, Massachusetts 02139
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  • Klavs F. Jensen

    Corresponding author
    1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Mass. Ave., 66-566, Cambridge, Massachusetts 02139; telephone: +1(617)253-4589; fax: +1(617)258-8224
    • Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Mass. Ave., 66-566, Cambridge, Massachusetts 02139; telephone: +1(617)253-4589; fax: +1(617)258-8224.
    Search for more papers by this author

Abstract

We describe a 150 µL microbioreactor fabricated in poly(methylmethacrylate) (PMMA) and poly(dimethylsiloxane) (PDMS) to cultivate microbial cell cultures. Mixing is achieved by a small magnetic stir bar and fluorescent sensors are integrated for on-line measurement of pH and dissolved oxygen. Optical transmission measurements are used for cell density. The body of the reactor is poly(methylmethacrylate) with a thin layer of poly (dimethylsiloxane) for aeration, oxygen diffuses through this gas-permeable membrane into the microbioreactor to support metabolism of bacterial cells. Mixing in the reactor is characterized by observation of mixing of dyes and computational fluid dynamics simulations. The oxygenation is described in terms of measured KLa values for microbioreactor, 20–75/h corresponding to increasing stirring speed 200–800 rpm. Escherichia coli cell growth in the microbioreactor is demonstrated and the growth behavior is benchmarked with conventional bench-scale bioreactors, flasks and tubes. Batch culture experiments with Saccharomyces cerevisiae further demonstrate the reproducibility and flexibility of the microbioreactor system. © 2005 Wiley Periodicals, Inc.

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