Tip from the experts. These parameters are adapted to water, if liquids of different properties and viscosities are used, these values need adjustment for reliable flow control.
Tip from the experts. For more details on the use of resistance to obtain the best performances in terms of flow rate control please refer to the “Flow control tuning” document.
4. Open an individually packed sterile syringe filter. Connect the 1/32” tubing to a male Luer fitting, then add a syringe membrane filter, and then add a female Luer fitting and length of 1/32” tubing.
Tip from the experts. Bacteria will tend to form biofilm inside all the tubing they have access to, which can affect the performance of the MFS2 and clog the resistance tubing over time. In order to protect the microfluidic circuit, a sterile syringe filter can be added inline to avoid bacteria reaching any upstream component and the sterile medium reservoir.
5. Connect the free end of the 1/32” tubing to a mini Luer fitting.
6. Under the laminar flow hood, remove the chip from its packaging, and inoculate it with the bacterial suspension. Use the pipette to completely fill the chip from one end to the other and close the inlets with parafilm to prevent contamination.
7. Connect the chip (since it is a straight channel chip, the inlet and outlet are interchangeable).
8. Connect 1/32” tubing to the chip outlet using a mini Luer fitting, and secure the other end to a safe waste container.
Tip from the experts.The waste contaminated with bacteria constitutes a biohazard. Thus, It must be contained according to the biosafety level of the stain used to ensure safety.
Tip from the experts. After incubation, air will be pushed first through the chip, displacing all the bacterial suspension. Most attached cells will remain attached after pushing air at the proposed flow rate. It is also an option to first purge air from the tubing with the chip disconnected, and reconnect the chip with liquid flowing at a slow drip.
2. Put the chip on the microscope stage. In the OB1 control window, choose the corresponding channel to the reservoir and flow the medium at 5 μl/min until the filter is almost filled with solution.
3. Set the flow rate value to 1 μl/min and let it run until the solution reaches the waste reservoir.
4. Leave the set up running for as long as needed. In this case, considerable biofilm growth was reached after 9 hours. Many biofilms can form in the same channel in one experiment, counting as replicates for the same conditions.
Application note written by Jesús Manuel Antúnez Domínguez
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