The purpose of the study was to measure nasal passageway dimensions in guinea pigs and rats by use of acoustic rhinometry (AR) and by a previously described fluid-displacement method (FDM) (Straszek SP, Taagehoej F, Graff S, and Pedersen OF. J Appl Physiol 95: 635-642, 2003) to investigate the potential of AR in pharmacological research with these animals. We measured the area-distance relationships by AR of nasal cavities postmortem in five guinea pigs (Duncan Hartley, 400 g) and five rats (Wistar, 250 g) by using custom-made equipment scaled for the purpose. Nosepieces were made from plastic pipette tips and either inserted into or glued onto the nostrils. We used liquid perfluorocarbon in the fluid-displacement study, and it was carried out subsequent to the acoustic measurements. We found for guinea pigs that AR measured a mean volume of 98 mm(3) (95-100 mm(3)) (mean and 95% confidence interval) of the first 2 cm of the cavity. FDM measured a mean volume of 146 mm(3) (117-175 mm(3)), meaning that AR only measured 70% (50-90) of the volume by FDM. For rats, the volume from 0 to 2 cm was 58 mm(3) (55-61 mm(3)) by AR and 73 mm(3) (60-87 mm(3)) by FDM, resulting in AR only measuring 83% (66-100%) of volume by FDM (see Table 2). We conclude that absolute nasal cavity dimensions are underestimated by AR in guinea pigs and rats. This does not preclude that relative changes may be correctly measured. In vivo trials with AR using rats have not yet been published. The FDM is possibly the most accurate alternative to AR for measurements of the nasal cavity geometry in small laboratory animals, but it can only be used postmortem.