Custom open-chamber microfluidic fabrication
Richard Wade-Martins, Quyen Do, Jimena Baleriola, Nora Bengoa-Vergniory
Abstract
This protocol described the fabrication of two- and three-open chambered microfluidics suitable for cell culturing using commercially acquired master mould.
Before start
Always store cut-out devices in ethanol.
Avoid exposing microfluidic devices, particularly the cell-adherent side, to open air and unnecessary contact. Once sterilised, devices should be handled with care following sterile working techniques.
Steps
Casting of microfluidic devices
A custom master mould containing both duo- and trio-chambers ( Figure 1 ) is manufactured by and obtained from Microliquid (Spain).
Mix 46 ml of silicon elastomer SylGARD 184, Dow Corning © with 4 mL of its curing agent (supplied in the same pack), i.e. the 1:10 ratio in a 50 ml falcon.
Mix well by inverting the falcon multiple times till a homogenous solution is seen by eye.
Slowly pour the mixture into the master mould and incubate for 2-2.5 hours at 60ºC.
Device preparation
Manually remove the final cast from the mould using a sharp blade.
Manually cut out individual devices and immediately wash them with 100% ethanol.
Air-dry cut-out devices in tissue culture hood.
Place dry and sterile devices on ethanol – sterilised 19 mm-diameter coverslips to form an instantaneously tight seal.
Coat both chambers and microchannel area with Poly-D-Lysin (0.1 mg/ml) overnight followed by Geltrex™ prior to cell culturing.
