Construction by dielectrophoresis of microbial aggregates for the study of bacterial cell dormancy |
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| Authors: | Ke Zhu Arseny S Kaprelyants Elena G Salina Martin Schuler and Gerard H Markx |
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| Institution: | 1School of Chemical and Analytical Science, The University of Manchester, Manchester M60 1Qd, United Kingdom;2Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow 117071, Russia;3Institute of Biochemical Engineering, University of Stuttgart, Germany;4Department of Chemical Engineering, Heriot-Watt University, Edinburgh, Scotland EH14 4AS, United Kingdom |
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| Abstract: | A study of the effect of aggregate size on the resuscitation of dormant M. smegmatis was conducted by constructing cell aggregates with defined sizes and shapes using dielectrophoresis and monitoring the resuscitation process under controlled laboratorial conditions in a long-term cell feeding system. Differently sized cell aggregates were created on the surface of indium tin oxide coated microelectrodes, their heights and shapes controlled by the strength of the induced electric field and the shape of the microelectrodes. Both two-dimensional (ring-patterned) and three-dimensional cell aggregates were produced. The cell aggregates were maintained under sterile conditions at 37 °C for up to 14 days by continuously flushing Sauton’s medium through the chamber. Resuscitation of dormant M. smegmatis was evaluated by the production of the fluorescent dye 5-cyano-2,3-ditolyltetrazolium chloride. The results confirm that the resuscitation of dormant M. smegmatis is triggered by the accumulation of a resuscitation promoting factor inside the aggregates by diffusion limitation. |
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