ibidi Pump System
Temperature changes along the tubing,
e.g., when parts of the tubing are led out-
side an incubator. This can happen when
the Perfusion System is set up directly on
the microscope.
Keep the temperature stable along the tubing, e.g.,
with a cage incubator with a temperature-controlled
atmosphere.
Temperature changes over a longer time
period, e.g., instabilities of the incubator
temperature control, or manifold open-
ings of the incubator door.
Keep the temperature stable. Avoid opening the incu-
bator door if not necessary.
10.2
Cells are Detaching
There are multiple parameters that influence cell attachment. Monitor the cells and make a note of
the time point from which the cells start to look unhealthy.
10.2.1
Cells Detach Before Starting the Flow
Issue:
The cells look unhealthy and do not attach to the slide surface before connecting them to the
Perfusion Set.
Possible Cause
Solution
The cells lack medium with nutrients. In
low channels (channel height 100 or 200
µm), the volume of medium is very low
with regard to the cell number.
Cultivate cells in low channels for only a few hours.
If the cells need to be in the channel for more than a
few hours, refresh the medium in the channel in short
intervals or place the slide on a rocking plate.
The cell culture surface is not suitable for
the specific cells.
Make sure the cells adhere to the surface under stan-
dard conditions (e.g. in a µ-Dish). If using a protein
coating, make sure the concentration of the coating
solution is sufficient for the channel slide. A protein
coating protocol is available in
The cells were not healthy.
Try another lot or passage of cells. Especially primary
cells are very variable in their fitness.
Evaporation in the slide led to increased
salt or glucose concentration, affecting
cellular viability.
Place the slide in an extra humidity chamber (Petri
dish with wet paper towel).
10.2.2
Cells Detach When Connecting the Slide to the Perfusion Set
Issue:
After connecting the Perfusion Set, the cells look detached or accumulated in clusters.
Possible Cause
Solution
The cells were detached by too much flow
during the connection step.
Every movement of the medium results in shear stress.
High flow rates can detach even healthy cells. Reduce
the flow by being more careful.
Instruction Manual
73
Version 2.6 (May 12, 2022)
