Automate cell seeding in a microfluidic chip for dynamic cell culture

Flow controller OB1 Mk3+

Mux-distributor

Microfluidic bubble trap

Flow sensor

Manifold 9 ports

Tubing, fittings and reservoirs

Automated cell seeding setup diagram

HARDWARE:

• Pressure & Flow controller: Impose a given pressure in order to create a stable and pulseless flow.
• Reservoirs: Contain your medium, buffer, stains, cell suspension or samples. Various cap sizes are available, from Eppendorf to bottles.
• Rotation valve (MUX-Distributor): Select the injected liquid.
• Flow Sensor: Monitor and control the flow rate in real time.
• Bubble trap: Trap and remove air bubbles to avoid them entering your chip, disturbing flow and adhered cells.
• Perfusion chamber or microfluidic chip: Contains your cells. Compatible with microscopy.
• Computer: Control all the parameters with our software and automate your experiment by creating injection sequences.

REAGENTS:

• Cells: MCF7 (1×10⁶ cells/mL)
• Medium: DMEM high glucose, 10% FBS, Penicillin/ Streptomycin (100 U/mL; 100 µg/mL)
• Wash buffer: PBS
• Live/ dead cell stains: Calcein/ Hoechst/ Propidium Iodide

Protocol for automated cell seeding

Assemble system: Connect all modules, calibrate the instrument, and prepare and connect all solutions.

Purge: Disconnect tubing from the inlet to the bubble trap. Purge all reservoir lines of air. Reconnect tubing.

Flow medium: Switch MUX-Distributor inlet to medium reservoir. Start flow until the perfusion chamber is filled with medium.

Flow cells: Switch MUX-Distributor inlet to cells and flow until perfusion chamber is filled with cells.

IMPORTANT: accurately count the cells and use a well-mixed suspension immediately.

Stop the flow: Switch MUX-Distributor inlet to dead-end to stop the flow.

TIP: It is helpful to monitor the channel under a microscope during seeding.

Incubate: Give the cells time to adhere to the surface of the chip while no liquid is flowing. This can be up to 24 h depending on cell type.

Results

Perfusion chamber for automated cell seeding at microscope stage.

TIP: chip can be secured in a Petri dish for easier handling.

MCF7 cells in chip, settling (t=2h).

MCF7 cells in chip, beginning to adhere (t=4h).

Seeding video: Cells are flowed into a perfusion chamber filled with medium at a flow rate of 40 µl/min. Flow was stopped by switching the MUX-distributor valve to a dead-end block at t=18 s, then stopping the OBI. Note the instantaneous arrest of cells.

Hints and tips

Conclusion

Automated cell seeding represents a major advancement in microfluidics, offering precision, reproducibility, and scalability for applications in tissue engineering, drug screening, and regenerative medicine. By minimizing human error and ensuring uniform cell distribution, automated systems enhance experimental reliability and accelerate research workflows.

For researchers seeking the most advanced and efficient microfluidic solutions, Elveflow provides state-of-the-art instruments and expertise to optimize automated cell seeding processes. With high-precision flow control systems, customizable setups, and dedicated technical support, Elveflow empowers scientists to achieve consistent and high-quality results in their cell culture experiments.

By integrating Elveflow’s cutting-edge technology into your lab, you can streamline your workflows, improve seeding accuracy, and unlock new possibilities in biomedical research. Trust Elveflow to deliver the performance and reliability needed for next-generation microfluidic applications.

Application note written by Lisa MUIZNIEKS, Subia BANO, Camila BETTERELLI GIULIANO and Jessica AYACHE.

Acknowledgement: This work was done thanks to the funding of European Union’s Horizon 2020 research and innovation programme (PANBioRA project, grant agreement No 760921; MECH-LoC project, MSCA grant agreement No 793749; MTOAC project, MSCA grant agreement No 795754; Protomet project, MSCA grant agreement No 813873).