Side-by-side comparison of disposable microchips with commercial capillary cartridges for application in capillary isoelectric focusing with whole column imaging detection.

Simple-structured, well-functioned disposable poly(dimethylsiloxane) (PDMS) microchips were developed for capillary isoelectric focusing with whole column imaging detection (CIEF-WCID). Side-by-side comparison of the developed microchips with well-established commercial capillary cartridges demonstrated that the disposable microchips have comparable performance as well as advantages such as absence of lens effect and possibility of high-aspect-ratio accompanied with a dramatic reduction in cost.

Integration of dialysis membranes into a poly(dimethylsiloxane) microfluidic chip for isoelectric focusing of proteins using whole-channel imaging detection.

A poly(dimethylsiloxane) microfluidic chip-based cartridge is developed and reported here for protein analysis using isoelectic focusing (IEF)-whole-channel imaging detection (WCID) technology. In this design, commercial dialysis membranes are integrated to separate electrolytes and samples and to reduce undesired pressure-driven flow. Fused-silica capillaries are also incorporated in this design for sample injection and channel surface preconditioning. This structure is equivalent to that of a commercial fused-silica capillary-based cartridge for adapting to an IEF analyzer (iCE280 analyzer) to perform IEF-WCID. The successful integration of dialysis membranes into a microfluidic chip significantly improves IEF repeatability by eliminating undesired pressure-driven hydrodynamics and also makes sample injection much easier than that using the first-generation chip as reported recently. In this study, two microfluidic chips with a 100-microm-high, 100-microm-wide and a 200-microm-high, 50-microm-wide microchannel, respectively, were applied for qualitative and quantitative analysis of proteins. The mixture containing six pI markers with a pH range of 3-10 was successfully separated using IEF-WCID. The pH gradient exhibited a good linearity by plotting the pI value versus peak position, and the correlation coefficient reached 0.9994 and 0.9995 separately for the two chips. The separation of more complicated human hemoglobin control sample containing HbA, HbF, HbS, and HbC was also achieved. Additionally, for the quantitative analysis, a good linearity of IEF peak value versus myoglobin concentration in the range of 20-100 microg/mL was obtained.

Method for microfluidic whole-chip temperature measurement using thin-film poly(dimethylsiloxane)/rhodamine B.

A novel method is presented for on-chip temperature measurements using a poly(dimethylsiloxane) (PDMS) thin film dissolved with Rhodamine B dye. This thin film is sandwiched between two glass substrates (one of which is 150 microm thick) and bonded to a microchannel molded in a PDMS substrate. Whole-chip (liquid and substrate) temperature measurements can be obtained via fluorescent intensity visualization. For verification purposes, the thin film was tested with a tapered microchannel subjected to Joule heating, with resulting axial temperature gradients comparing well with numerical simulations. Errors induced by the definite film thickness are discussed and accounted for during experimental and analytical analysis. Alternative validation using the traditional in-channel Rhodamine B injection method was also attempted. The thin film has several advantages over traditional methods. First, false intensity readings due to adsorption and absorption of Rhodamine B into PDMS channels are eliminated. Second, whole-chip temperature measurements are possible. Third, separation of working liquid from Rhodamine B dye prevents possible electrophoresis effects.