Automated recirculation perfusion system
Continuous, unidirectional flow over samples
No need to switch between reservoirs
The cell culture pump replaces the need for a CO2 incubator
No more losing your experiment due to clogging
Automated recirculation perfusion system
Our automated recirculation perfusion system is designed to maintain a stable and controlled environment for long-term cell culture. The continuous recirculation allows for real-time monitoring and provides more accurate data on the experimental conditions. Moreover, the recirculation perfusion system reduces the needed reagents compared to single-pass systems, giving you even more control over your experiments.
Moreover, it ensures continuous nutrient supply at constant O2 and CO2 levels without the need for a CO2 incubator. This precision also provides a more physiologically relevant cell microenvironment by sustaining a continuous unidirectional flow through a microfluidic chip laden with cells.
From reservoir 1 to reservoir 2:
From reservoir 2 to reservoir 1:
Automated recirculation perfusion system applications
The automated recirculation perfusion system setup includes:
- Level sensors
- Cell culture pump
- Check valves (recirculation bridge)
- Microfluidic flow sensor (eg. Galileo)
- Several Schott bottles
- Tubings and fittings
- Microfluidic chip
- User guide
- Software (Galileo user interface)
Some biologial applications of our recirculation perfusion system include:
Have a look at this review comparing this unidirectional automated recirculation system to unidirectional recirculation with a peristaltic pump.
Check this review comparing different bidirectional and unidirectional recirculation systems.
With microfluidics it is possible to flow medium continuously over cells in culture. A microfluidic recirculation perfusion system involves fluid’s continuous and controlled circulation through microchannels.
Recirculation perfusion systems
Most recirculation perfusion systems rely on the initial fluid volume and desired flow rates to estimate the time it takes for one reservoir to flow most of its volume through the sample before switching to the other reservoir.
However, these systems don’t detect changes and thus continue functioning as programmed. If one of the reservoirs empties before expected due to biofouling or clogging, air enters the system, damaging the cells and ruining the experiment.
In addition, a recirculation perfusion system requires the medium to be supplied with CO2 inside the chip. This can be done using a gas-permeable material, a CO2-independent medium for short periods (about 2 hours), or a CO2 incubator.
One pack for an automated recirculation perfusion system
We have combined the cell culture pump, check valves, and level sensors in one pack for automated recirculation perfusion.
The cell culture pump can be used in the setup as a pressure controller that consumes less gas than other pressure-driven controllers. Pressuring the media reservoir with the correct gas mix prevents the gas in the media from diffusing into the atmospheric air.
The constant media flow ensures that the correct composition always reaches the cells, even if the rest of the system is not gas-tight.
The cell culture pump allows recirculation outside of a CO2 incubator. In this case, an additional stage-top incubator might be used to keep the temperature stable for long periods of time, allowing for high-quality live cell imaging experiments.
The check valve recirculation bridge keeps the flow unidirectional inside the connected microfluidic chip via four passive check valves that are adjusted to quickly and efficiently transfer fluid between reservoirs.
In the figure below, the small red arrows indicate the direction of flow inside each check valve. Corner number 1 marks the common exit from the bridge.
Check this application note for more information on how to assemble the recirculation bridge and set up your experiment.
The level sensors automate the perfusion system based on the media volume level inside reservoirs, ensuring continuous perfusion. In addition, the level sensors introduce a fail-safe mechanism that considers the ever-evolving nature of a perfusion system, such as fouling the tubing with dead cell debris, leakage due to clogging, and other blockage-related issues, such as air entry.
For example, in case of leakage due to extensive clogging, the level sensors will stop the flow, preventing air from being pushed into the system and preserving the sample.
A highly-performant flow sensor for clogging detection
Our engineers have developed a new highly-performant flow sensor, Galileo. This sensor offers <5% flow rate accuracy for sensing ranges from 0.5 to 10,000 µL/min.
Galileo allows for bi-directional flow rate measurement and automatic detection of internal clogging, which will be shown on the small screen of the flow sensor. In case of cross-contaminations or clogging, it’s possible to replace the cartridge for an entirely new flow path. The Galileo flow sensor is a key component that ensures the system’s accuracy and reliability.
Check the whole video here.
Level sensor technical specifications
The following table summarizes the main specifications of the level sensors:
Components | Technical specifications |
Dimensions (cm) | Sensing area: 2 x 1 cm |
Material | Plastic |
Reservoir compatibility | 100 mL Schott bottle |
Control | Cell culture pump |
Cell culture pump technical specifications
The cell culture pump can come with 1 to 4 channels. Each channel connects to a flow sensor with a feedback loop between pressure and flow rate. Each channel has the following specifications (tested with a set pressure value of 2 bar):
Characteristics | Specifications |
Accuracy | -27.75 mbar |
Air consumption | 0.24 L/min |
Response time | 140 ms |
Settling time | 2750 ms |
Overshoot | 0.12 mbar |
Customize your pack
Our instruments can benefit from your feedback, so you can take advantage of the extra flexibility to adapt them according to your specific needs. Our microfluidic specialists will advise you on the best instruments and accessories based on your needs and will accompany you during the setup of the recirculation perfusion system.
All the instruments are controlled by the same software, allowing workflow automation and easy integration in your program with free available libraries.
Is the recirculation perfusion system compatible with reservoirs other than bottles?
The current version is compatible with 100 mL bottles, but other options, such as for well-plates, are being developed. Don’t hesitate to contact us if you need a different reservoir type.
Does the cell culture pump keep the gas sterile?
To guarantee the sterility of the used gases, we advise adding a small disposable filter at the gas inlet of the reservoir.
Which microfluidic chips can be connected to the cell culture pump?
The cell culture pump can be connected to any microfluidic chip using the correct connectors.
Can the cell culture pump be placed inside the CO2 incubator?
The cell culture pump is intended to replace the need for a CO2 incubator.
What is the maximum flow rate that can be applied?
The system works well with the range of 0-5ml/min.
Funding and Support
The LIFESAVER project, funded by the European Union’s H2020-LC-GD-2020-3, grant agreement No. 101036702 (LIFESAVER), helped develop the level sensors.
The Tumor-LN-oC project, funded by the European Union’s H2020-NMBP-TR-IND-2020 grant agreement No. 953234 (Tumor-LN-oC), helped develop the cell culture pump.
The ALTERNATIVE project, funded by the European Union’s H2020-LC-GD-2020-3 grant agreement No. 101037090, helped develop the check valve system.
The Galileo project, funded by the European Union’s Horizon research and innovation program under HORIZON-EIC-2022-TRANSITION-01 grant agreement No. 101113098 (GALILEO), helped develop the Galileo flow sensor.
