Pressure pulse injection: a powerful alternative to electrokinetic sample loading in electrophoresis microchips

We present a new and simple capillary electrophoresis microsystem in which the sample is injected hydrodynamically using a pressure pulse. This technique maintains the sample composition, in contrast to a classical electrokinetic injection, in which the magnitude of the electric field in the sample reservoir in combination with variations in electrophoretic mobility can lead to a biased injection. The sample is loaded using a well controlled and variable pressure pulse (0.1-1.0 s) generated by the mechanical actuation of a flexible membrane placed on the chip sample reservoir. A fluorescein/calcein-containing borate-Tris-hydroxymethylaminoethane (TRIS) sample solution is taken as a model system for CE analysis. The separation results using pressure pulse injection clearly demonstrate the advantages of our technique. In addition to the reduced bias due to the absence of an electrode in the sample well, this method allows injection of variable plug volumes by simply changing the pulse length. Moreover, a very high speed, repeatability, and sensitivity of the separation is obtained.