Monitoring Cellular Stress Responses using Integrated High-Frequency Impedance Spectroscopy and Time-Resolved ELISA

Verena Charwat, Martin Joksch, Drago Sticker, Michaela Purtscher, Mario Rothbauer, Peter Ertl

    Publikation: Beitrag in FachzeitschriftArtikelBegutachtung

    Abstract

    We have developed a lab-on-a-chip system for continuous and non-invasive monitoring of microfluidic cell cultures using integrated high-frequency contactless impedance spectroscopy. Electrically insulated microfabricated interdigitated electrode structures were embedded into four individually addressable microchambers to reliably and reproducibly detect cell-substrate interactions, cell viability and metabolic activity. While silicon nitride passivated sensor substrates provided a homogeneous cell culture surface that minimized cell orientation along interdigitated electrode structures, the application of highfrequency AC fields reduced the impact of the 300 nm thick passivation layer on sensor sensitivity. The additional implementation of multivariate data analysis methods such as partial least square (PLS) for high-frequency impedance spectra provided unambiguous information on intracellular pathway activation, up and down-regulation of protein synthesis as well as global cellular stress responses. A comparative cell analysis using connective tissue fibroblasts showed that high-frequency contactless impedance spectroscopy and time-resolved quantification of IL-6 secretion using ELISA provided similar results following stimulation with circulating pro-inflammatory cytokines IL-1b and TNFa. The combination of microfluidics with contactless impedance sensing and time-resolved quantification of stress factor release will provide biologist with a new tool to (a) establish a variety of uniform cell culture surfaces that feature complex biochemistries, micro- and nanopatterns; and (b) to simultaneously characterize cell responses under physiologically relevant conditions using a complementary noninvasive cell analysis method.
    OriginalspracheEnglisch
    Seiten (von - bis)5271-5282
    Seitenumfang12
    FachzeitschriftAnalyst
    Volume139
    Issue20
    DOIs
    PublikationsstatusVeröffentlicht - 2014

    Research Field

    • Biosensor Technologies

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