65- Place the vessel on top of the stirrer and on the heat pad.
- Connect the probes to the correct channel on the Eycoferm
controller, which will maintain growth parameters at set values
within the culture vessel. - Check the oxygen probe has a signal and a rough span between
0% dissolved air saturation (DAT) (via nitrogen addition) and
100% DAT (via air addition) (see Note 5). - Calibrate the pH probe with pH 4.0 and pH 7.0 buffers. To
control the pH (which is optional), the acid (1 M HCl) should
be delivered in response to a fluctuation in acidity levels via
peristaltic pumps, which will also respond to the Eycoferm
controller via a negative feedback loop (see Note 6). - Heat the water in the vessel to 37 °C using the heat pad. To
calibrate the temperature probe use a Tempcon hand-held dig-
ital thermometer to monitor the temperature in the vessel until
it reaches 37 °C. Set the temperature reading on the Eycoferm
controller to 37 °C (see Note 7).
Fig. 1 Description of diagram labels: (1) Pipette assembly for flow rate calibration, (2) Media line pump, (3)
Media reservoir, (4) Sample port assembly, (5) Effluent reservoirs, (6) Effluent line pump, (7) Chemostat vessel,
(8) Magnetic stirrer disc, (9) Heat mat, (10) Magnetic stirrer, (11) Anti-grow-back device on media line, (12) Air
Inlet assembly, (13) Air outlet/Condense bottle assembly, (14) DOT probe, (15) Temperature probe, (16) pH
probe, (17) Eycoferm controller, (18) Acid addition pump controlled by feedback loop from pH probe, (19)
Temperature control feedback loop, (20) DOT control feedback loop controlling stirrer speed
Application of Continuous Culture for Assessing Antibiotic Activity Against...