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Accurate size profiling on the physical sizes of aqueous nanoparticles and molecules may be affected by various factors such as the particle aggregation or the small molecular sizes. Thus, a nanoparticle exposure chamber system was constructed in which the aqueous nanoparticles are electrosprayed and dispersed into a controllable manner prior to be determined on their size distribution and concentration. The sizes determined by our system on polystyrene nanobeads are in good accordance with dynamic light scattering (DLS) and TEM. The system was further used to measure the physical sizes of bovine serum albumin (BSA), blue dextran and hybrid nanoparticles whose sizes are not easily measured with either DLS or TEM. In addition to serving as a complementary tool on the size and concentration measurements of nanoparticles, this system can also be used as a quantitative exposure protocol for airborne nanoparticles with defined nanoparticle sizes and concentrations.