Electrokinetic stacking of particle zones in confined channels enabling their UV absorbance detection on microchips

In this article, we report a simple approach to stacking micro- and nanoparticle zones by electrokinetically migrating them through moderately confined channels of uniform cross-section. Experiments show the reported pre-concentration process to initiate at the tail end of the zone following its electrokinetic injection, with the stacked region migrating faster than the rest of the sample band. This effect causes the particles traveling in front to merge into the stacked region making it grow both in size and concentration. Because the stacked zone also gradually loses particles from its trailing edge, it eventually disintegrates upon running out of particles at its front end. Nevertheless, enhancements in peak height by over 100-fold were recorded using the reported approach for polystyrene beads with diameters comparable to the channel depth. This enhancement however, exhibited a temporal variation as the particle band migrated through the analysis column reaching a maximum value that depended on the particle diameter, particle concentration, channel depth, electric field strength, electroosmotic mobility, etc. Interestingly, the peak area recorded by the detector remained relatively constant during this particle migration period allowing reliable sample quantitation. Moreover, upon incubating antibody-coated particles against an antigen sample, the peak area for the particle zone was seen to scale linearly with the antigen concentration establishing the utility of the reported focusing phenomenon for chemical/biochemical analysis. The noted stacking technique was further applied to enabling UV absorbance detection of particle zones on microchips which then allowed us to determine the colloidal content in actual natural water samples. .