Automatic flow in fluid-walled dumbbells driven by Laplace pressure
Richard Wade-Martins, Quyen Do, Federico Nebuloni
Abstract
This protocol describes experiments performed to quantify pressure and volume variations inside fluid-walled dumbbells when a pressure difference is generated between the two chambers. Difference in Laplace pressure automatically flows medium from the high-pressure chamber to the low pressure one. Flow stops when pressures are equilibrated.
Steps
Jet-Printing of Fluid-Walled Dumbbells
Fill a virgin uniwell plate with 5 ml of DMEM supplemented with 10% FBS. Agitate the plate to spread the 5 mL of medium to cover the whole area of the plate.
Remove the volume leaving a thin layer of medium wetting the plate.
Gently overlay FC40 (~50 mL) pouring it in one of the corners.
Place the plate in the 3D traverse. Adjust x, y, z positions of the traverse head to coincide with the starting point of the movement path.
Start the pump, setting a constant flow at 480 μL/min.
Wait ~10 s to allow flow to be fully developed.
Start traverse automatic movement.
At completion of the movement path, stop pump and remove plate from the traverse.
Establishment of Pressure Difference and Volumes Variation Measurements
Place the newly printed plate into the pendant drop tensiometry machine.
Add FC40 to almost fill the plate entirely (~20 mL).
By mean of a syringe pump connected to a needle through a Teflon tube, infuse 4 μL in the right chamber and soon after 1 μL in the left chamber (infusion rate set on pump = 20 μl/min).
Record chambers height for 24 hours imaging every 30 minutes.
