Band Deformation at a T-Junction While Electrofocusing in a Dog-Leg Microchannel

On-chip isoelectric focusing (IEF) has been performed in both straight and dog-leg microchannels. Three-dimensional microfluidic chips were fabricated on poly di-methyl-siloxane (PDMS) using soft lithography and multilayer bonding techniques. Plasma oxidized PDMS channel surfaces were dynamically coated with methyl cellulose to discourage electroosmotic flow during separation and purification processes. In a straight microchannel, IEF was completed within 5 minutes at an applied electric field strength of 50 V/cm using broad range ampholytes. The focused bands were generally well-formed with sharp edges and were less than 100 microns across yielding a putative peak capacity in excess of 100 peaks in a 2-cm long channel. However, the conventional IEF protocol shifts the focused bands toward the cathodic well. This cathodic drift can be effectively minimized by placing highly viscous polymer solutions in the electrode reservoirs. In dog-leg microchannels, initially well formed focused band dispersed at the Tee-channel junction, but refocused at the dog-leg channels with relatively lower resolution.