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We present a new approach for fabricating multilayer microfluidic devices in poly(methyl methacrylate). Paraffin wax was used as a phase-changing sacrificial layer to protect microstructures during solvent bonding. Microchannels in the top and bottom pieces were aligned with through-holes in the middle layer, resulting in microchannels that cross one another. No discernible delamination of the layers or leakage between channels was observed at pressures as high as 300 psi. The current versus voltage linearity in the crossover channel indicates that no Joule heating occurs at voltages of at least 2.0 kV. Moreover, a potential in the crossover channel did not affect the current in the separation channel. Rapid and efficient separation of fluorescently labeled amino acids was performed in these devices. Pressurized buffer flow or voltage applied in the crossover channel caused no leakage into or electrical interference with the separation channel. Our results demonstrate that sacrificial layers with solvent bonding can be implemented readily in the fabrication of robust and fluidically complex multilayer polymer microchips. These capabilities should facilitate the development of a new generation of sophisticated microfluidic systems. [ABSTRACT FROM AUTHOR]