A fully integrated isotachophoresis with a programmable microfluidic platform.

Conventional isotachophoresis (ITP) can be used for pre-concentration of a single analyte, but preconcentration of multiple analytes is time consuming due to handling and washing steps required for the extensive buffer optimization procedure. In this work, we present a programmable microfluidic platform (PMP) to demonstrate fully automated optimization of ITP of multiple analytes. By interfacing a PMP with ITP, buffer selection and repetitive ITP procedures were automated. Using lifting-gate microvalve technology, a PMP consisting of a two-dimensional microvalve array was designed and fabricated for seamless integration with an ITP chip. The microvalve array was used for basic liquid manipulation such as metering, mixing, selecting, delivering, and washing procedures to prime and run ITP. Initially, the performances of the PMP and ITP channel were validated individually by estimating volume per pumping cycle and preconcentrating Alexa Fluor 594 with appropriate trailing (TE) and leading (LE) buffers, respectively. After confirming basic functions, autonomous ITP was demonstrated using multiple analytes (Pacific blue, Alexa Fluor 594, and Alexa Fluor 488). The optimal buffer combination was was determined by performing multiple ITP runs with three different TEs (borate, HEPES, and phosphate buffers) and three different concentrations of Tris-HCl for the LE. We found that 40 mM borate and 100 mM Tris-HCl successfully preconcentrated all analytes during a single ITP run. The integrated PMP-ITP system can simplify overall buffer selection and validation procedures for various biological and chemical target samples. Furthermore, by incorporating analytical tools that interconnect with the PMP, it can provide high sample concentrations to aid in downstream analysis.