ibidi Pump System
6.7
Connect the Cell-Seeded Slide to the Perfusion Set
Note!
After the calibration, remove the cell-free slide and re-connect the female Luer middle connector.
Once again remove the air bubbles and then connect the slide with the cells.
Three points are imperative when connecting the µ-Slide to the Perfusion Set:
• Avoid air bubbles, which can remove seeded cells from the slide.
• Maintain sterility by proper sterile handling while seeding and pre-cultivating cells, as well as
filling the reservoirs.
• Avoid disturbance of the cells, such as strong temperature variations or fast flow pulses.
The connection procedure is the same as shown in Section
. For a detailed description, refer to
Application Note 13, “Endothelial Cells Under Perfusion”
6.8
Start the Flow
After connecting the slide to the tubing, check the cells under the microscope. It is crucial that the
cell layer is confluent and well adherent when it is exposed to shear stress. If the cells are stressed it
may be better to let them recover before initiating flow.
To start flow, place the whole assembly (Fluidic Unit and slide) in the incubator and connect the
Fluidic Unit to the pump (air pressure tubing and electric cable). Start the experiment in the ibidi
Pump Control software by increasing the shear stress step by step until the target shear stress is
reached (see
and
6.9
Observing Cells on the Microscope
To observe your cells on the microscope stop the pump when the levels in the reservoirs are equili-
brated. Then detach the air pressure tubing and the electric cable of the Fluidic Unit. Take the Fluidic
Unit with the connected µ-Slide to the microscope, put it beside the specimen holder, and place the
slide into the holder.
After the observation, place the Fluidic Unit and slide back in the incubator and restart the flow.
Note!
This procedure does not affect sterility! Take care not to disconnect any adapter or the filters on
top of the Fluidic Unit reservoirs!
Make sure that the Fluidic Unit does not stay longer than 10 minutes outside the incubator to
avoid cooling effects, such as stress on the cells or air bubble generation!
Version 2.6 (May 12, 2022)
48
Instruction Manual
