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Detecting Bacterial Cell Viability in Few µL Solutions from Impedance Measurements on Silicon-Based Biochips
Authors:Vinayak J. Bhat,Sahitya V. Vegesna,Mahdi Kiani,Xianyue Zhao,Daniel Blaschke,Nan Du,Manja Vogel,Sindy Kluge,Johannes Raff,Uwe Hü  bner,Ilona Skorupa,Lars Rebohle,Heidemarie Schmidt
Affiliation:1.Leibniz Institute of Photonic Technology, Albert-Einstein-Str. 9, 07745 Jena, Germany; (V.J.B.); (D.B.); (U.H.);2.Center for Microtechnologies, Chemnitz University of Technology, 09126 Chemnitz, Germany; (M.K.); (X.Z.);3.Institute for Solid State Physics, Friedrich Schiller University Jena, Helmholtzweg 3, 07743 Jena, Germany;4.Helmholtz-Zentrum Dresden-Rossendorf, Bautzner Landstraße 400, 01328 Dresden, Germany; (M.V.); (S.K.); (J.R.); (I.S.); (L.R.)
Abstract:Using two different types of impedance biochips (PS5 and BS5) with ring top electrodes, a distinct change of measured impedance has been detected after adding 1–5 µL (with dead or live Gram-positive Lysinibacillus sphaericus JG-A12 cells to 20 µL DI water inside the ring top electrode. We relate observed change of measured impedance to change of membrane potential of L. sphaericus JG-A12 cells. In contrast to impedance measurements, optical density (OD) measurements cannot be used to distinguish between dead and live cells. Dead L. sphaericus JG-A12 cells have been obtained by adding 0.02 mg/mL of the antibiotics tetracycline and 0.1 mg/mL chloramphenicol to a batch with OD0.5 and by incubation for 24 h, 30 °C, 120 rpm in the dark. For impedance measurements, we have used batches with a cell density of 25.5 × 108 cells/mL (OD8.5) and 270.0 × 108 cells/mL (OD90.0). The impedance biochip PS5 can be used to detect the more resistive and less capacitive live L. sphaericus JG-A12 cells. Also, the impedance biochip BS5 can be used to detect the less resistive and more capacitive dead L. sphaericus JG-A12 cells. An outlook on the application of the impedance biochips for high-throughput drug screening, e.g., against multi-drug-resistant Gram-positive bacteria, is given.
Keywords:cell viability   Lysinibacillus sphaericus   impedance biochips   membrane potential   dead bacterial cells   live bacterial cells
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