Pneumatic equiaxial compression device for mechanical manipulation of epithelial cell packing and physiology
Fig 2
The device was set inside the mini-incubator and connected to the vacuum pressure controller. The vacuum battery was attached between the device and the vacuum pressure controller. During the cell culture period the mini-incubator containing the device was kept inside a standard cell culture incubator. The valve of the vacuum battery was opened, thus allowing the vacuum pressure controller to maintain pressure during cell culture. After the six-day cell culture period during transportation to the microscope, the valve of the vacuum battery was closed, which allows the setup to be momentarily detached from the vacuum pressure controller as the vacuum battery maintains the vacuum pressure. At the microscope, the device was again connected to the vacuum pressure controller and the valve was opened. Gas flow (5% CO2) to the mini-incubator was initiated once the device was removed from the standard incubator to maintain pH during transportation and imaging. Heated gas to maintain the temperature during the imaging was supplied into the microscope stage top incubator.