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Issue 20, 2014
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Monitoring cellular stress responses using integrated high-frequency impedance spectroscopy and time-resolved ELISA

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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 high-frequency 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-1β and TNFα. 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 non-invasive cell analysis method.

Graphical abstract: Monitoring cellular stress responses using integrated high-frequency impedance spectroscopy and time-resolved ELISA

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Publication details

The article was received on 07 May 2014, accepted on 11 Aug 2014 and first published on 11 Aug 2014


Article type: Paper
DOI: 10.1039/C4AN00824C
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Citation: Analyst, 2014,139, 5271-5282

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    Monitoring cellular stress responses using integrated high-frequency impedance spectroscopy and time-resolved ELISA

    V. Charwat, M. Joksch, D. Sticker, M. Purtscher, M. Rothbauer and P. Ertl, Analyst, 2014, 139, 5271
    DOI: 10.1039/C4AN00824C

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