Accurately determining the delivered dose is critical to understanding biological response due to cell exposure to chemical constituents in aerosols. Deposition efficiency and uniformity of deposition was measured experimentally using monodisperse solid fluorescent particles with mass median aerodynamic diameters (MMAD) of 0.51, 1.1, 2.2 and 3.3 μm in the Vitrocell® AMES 48 air-liquid-interface (ALI) in vitro exposure system. Experimental results were compared with computational fluid dynamic, (CFD; using both Lagrangian and Eulerian approaches) predicted deposition efficiency and uniformity for a single row (N = 6) of petri dishes in the Vitrocell® AMES 48 system. The average experimentally measured deposition efficiency ranged from 0.007% to 0.43% for 0.51–3.3 μm MMAD particles, respectively. There was good agreement between average experimentally measured and the CFD predicted particle deposition efficiency, regardless of approach. Experimentally measured and CFD predicted average uniformity of deposition was greater than 45% of the mean for all particle diameters. During this work a new design was introduced by the manufacturer and evaluated using Lagragian CFD. Lagragian CFD predictions showed better uniformity of deposition, but reduced deposition efficiency with the new design. Deposition efficiency and variability in particle deposition across petri dishes for solid particles should be considered when designing exposure regimens using the Vitrocell® AMES 48 ALI in vitro exposure system.