In this study, we report development of novel compartmental microfluidic chip devices for multiplexed single-cell sorting and analysis of antigen-specific single B-cells before and after stimulation with an antigen from a bulk cell suspension in the aqueous compartments that are generated by the two-phase liquid systems in the Y-shaped microchannels made from poly (dimethylsiloxane) (PDMS) polymer. Microfabrication technology allowed us to build the microfluidic device, which established the stable and high-throughput picoliter compartments of two-phase liquids under pressure-driven force due to viscosity and interfacial tension of liquids. Optimization of compartmental microfluidics revealed they could generate over ~30 compartments per sec with a size of < 150 pl volume suggest they are best suitable for single molecule and single-cell assays. This chip system has the ability of sorting up to ~1250 cells <100 sec. These devices are best suitable for high-sensitive on-chip bioassays since rapid mixing is possible in the compartments, hence this system serve as a micro total analysis system in diagnostic applications etc.