Diagram of AIO interdigitated castellated electrodes with characteristic dimensions from 25 to .
Design of the ring-patterned microelectrode for the construction of 2D aggregates.
Closed feeding chamber built on the top of an AIO microelectrode fitted with an injection port.
Illustration of the different aggregate shapes formed. A concave aggregate would have a ratio of aggregate height at the edge of the electrode to that at the center that is larger than 1.
Procedures used for patterning and immobilizing two strains of microorganisms in a 2D pattern. (a) Energized electrodes before introduction of first cell type. (b) Cells are attracted to high field regions and accumulate in outer rings after their introduction; cells are then immobilized. (c) On energizing the two inner electrodes and grounding the outer electrodes, cells of the second cell type preferentially accumulate at the center; cells are then immobilized.
Diagram of the system used for studying the resuscitation of dormant M. smegmatis. Fresh Sauton’s medium was pumped into the chamber at a flow rate of for 8–12 days; the chamber was kept at and resuscitation was checked daily. (a) Feed tank with Sauton’s medium; (b) air filter; (c) microelectrode chamber with cell aggregates; (d) waste collection.
Aggregates of M. smegmatis w-t cells in an AIO chamber with electrodes with characteristic sizes ranging from 25 to . The applied signal was 1 MHz, . Dark areas are formed by cells; the arrows point at the ITO electrodes.
Close-up of aggregates of active M. smegmatis w-t cells. The applied signal was 1 MHz, .; aggregates are shown 30 min after the start of the aggregation process. Darker areas indicate the presence of cells; the arrows point at the ITO electrodes. (a) Typical biofilm with overlapping small aggregates in the region. (b) Single aggregate formed in the region.
The height of M. smegmatis w-t aggregates at different characteristic sizes of the microelectrodes. The applied electric signal was 1 MHz, . (a) Height at the edges of the castellations, 45 min after the start of the aggregation process; (b) height at the center of the aggregates, 45 min after the start of the aggregation process.
The height of M. smegmatis w-t aggregates at different characteristic sizes of the microelectrodes at the center of the aggregates, 6 h after the start of the aggregation process. The applied electric signal was 1 MHz, .
Resuscitation of dormant M. smegmatis (pAGR). (a) Composite picture showing fluorescent images of formazan formation; (b) average fluorescence intensity per aggregate as a function of incubation time.
Resuscitation of a 50:1 mixture of dormant M. smegmatis w-t and active M. luteus. (a) Composite picture showing fluorescent images of the formazan formation; (b) average fluorescence intensity per aggregate as a function of the incubation time.
Average fluorescence intensity per aggregate at day 4 of dormant M. smegmatis (pAGR) and of a 50:1 mixture of dormant M. smegmatis w-t mixed with active M. luteus as a function of aggregate size.
Bright field images taken during the construction 2D aggregates consisting of dormant M. smegmatis w-t and active M. luteus cells. (a) Dormant M. smegmatis were immobilized in the outer rings; the center of the structure was left empty; (b) same system after immobilization of M. luteus in the center.
Assessment of the resuscitation of M. smegmatis over time in a 2D system with no diffusion limitation. (a) Fluorescence image of system after the first day, (b) fourth day, and (c) fifteenth day. (d) Average intensity for the two different regions in the aggregate as a function of the incubation time.
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