on the cartridge and purge excess volume from the RH syringe
through the ECS and into the CR/duet circulation. Close the
RH ECS port.
9. With the ECS ports on the HF cartridge closed, detach syringes while holding a disinfectant wipe around the syringe/
microclave connection. Discard syringes into a biohazard
sharp box containing 20% bleach solution. Clean the HFC
ports, external surfaces of the cartridge and associated tubing
with a disinfectant wipe.
10. Return the CR and cartridge assembly to the 37
C incubator
and resume the flow of medium through the system by turning
on the peristaltic pump, opening the stopcock primary flow
path valve and releasing any tubing clamps.
3.8 Sampling from
the ECS
1. Halt the flow of medium by turning off the peristaltic pump
and close the stopcock valve above the CR. Remove the cassette
from the duet pump and close the terminal ports on the bioreactor. Move the cartridge and attached tubing to the BSC.
2. Wipe the external surfaces of the bioreactor and its ECS ports
with a disinfectant wipe followed by an alcohol swab to avoid
external contamination while sampling from the ECS. Close
the terminal ports on the cartridge and attach a sterile 3 mL
Luer-lock syringe to the microclave on the LH ECS port of the
cartridge.
3. Open the LH terminal port and the RH ECS port and draw
medium slowly into the RH syringe. Any bubbles that have
accumulated in the bioreactor may also be cleared in this
manner.
4. Close the LH terminal port and open the LH ECS port.
5. Use the extra medium in the RH syringe to mix the culture
back and forth between the two syringes to distribute the
culture evenly.
6. After multiple exchanges, push the culture volume in the LH
syringe through to the RH syringe without introducing air into
the cartridge and then close the LH ECS port.
7. Open the RH terminal port and push excess medium in the RH
syringe gently out of the cartridge and into the duet tubing
until the RH syringe is emptied.
8. Close the RH ECS port.
9. Open the LH ECS port and draw a sample of culture (typically
100–200μL) into the syringe.
10. Close the port and then detach the sample-containing syringe
while holding a disinfectant wipe around the microclavesyringe junction to contain any potential aerosolized culture.
724
A. Brett Mason and Ve ´ ronique Dartois
through the ECS and into the CR/duet circulation. Close the
RH ECS port.
9. With the ECS ports on the HF cartridge closed, detach syringes while holding a disinfectant wipe around the syringe/
microclave connection. Discard syringes into a biohazard
sharp box containing 20% bleach solution. Clean the HFC
ports, external surfaces of the cartridge and associated tubing
with a disinfectant wipe.
10. Return the CR and cartridge assembly to the 37
C incubator
and resume the flow of medium through the system by turning
on the peristaltic pump, opening the stopcock primary flow
path valve and releasing any tubing clamps.
3.8 Sampling from
the ECS
1. Halt the flow of medium by turning off the peristaltic pump
and close the stopcock valve above the CR. Remove the cassette
from the duet pump and close the terminal ports on the bioreactor. Move the cartridge and attached tubing to the BSC.
2. Wipe the external surfaces of the bioreactor and its ECS ports
with a disinfectant wipe followed by an alcohol swab to avoid
external contamination while sampling from the ECS. Close
the terminal ports on the cartridge and attach a sterile 3 mL
Luer-lock syringe to the microclave on the LH ECS port of the
cartridge.
3. Open the LH terminal port and the RH ECS port and draw
medium slowly into the RH syringe. Any bubbles that have
accumulated in the bioreactor may also be cleared in this
manner.
4. Close the LH terminal port and open the LH ECS port.
5. Use the extra medium in the RH syringe to mix the culture
back and forth between the two syringes to distribute the
culture evenly.
6. After multiple exchanges, push the culture volume in the LH
syringe through to the RH syringe without introducing air into
the cartridge and then close the LH ECS port.
7. Open the RH terminal port and push excess medium in the RH
syringe gently out of the cartridge and into the duet tubing
until the RH syringe is emptied.
8. Close the RH ECS port.
9. Open the LH ECS port and draw a sample of culture (typically
100–200μL) into the syringe.
10. Close the port and then detach the sample-containing syringe
while holding a disinfectant wipe around the microclavesyringe junction to contain any potential aerosolized culture.
724
A. Brett Mason and Ve ´ ronique Dartois