Fabrication of biofunctionalized microfluidic structures by low-temperature wax bonding.

In this work, a new fabrication technology for microfluidics based on the use of wax is described. Microfluidic structures are assembled using wax as both a thermoplastic adhesive layer between two glass substrates and a spacer layer defining the microchannels. Wax patterns with dimensions down to 25 μm are easily produced on glass substrates using specially developed decal-transfer microlithography. A complete microfluidic system is created by bonding the wax patterned layer with an additional glass substrate. On the basis of the special melting behavior of waxes, an effective glass-wax bonding is achieved at 40 °C by applying a soft pressure and without the requirement of any glass pretreatment. Wax bonding provides an effective sealing of the fluidic networks even on nonflat glass substrates (i.e., containing metal electrodes). The mild conditions required for the bonding process enables the fabrication of lab-on-a-chip devices incorporating biomolecules, as is demonstrated with the implementation of a simple heterogeneous immunoassay in a microfluidic device with amperometric detection.