This study assessed the collection efficiency (CE) of two popularly used sampling devices (BioSampler and Coriolis sampler) for fungal aerosols. Phosphate‐buffered saline (PBS) supplemented with or without surfactant (Tween‐20, Tween‐80, or Triton X‐100) and antifoam agent was prepared and used as collection liquids. The agar impactor (BioStage) was simultaneously operated with liquid‐based samplers to collect fungi from seven sites located at a university building, public library, and animal farming. Fungal concentrations determined by liquid samplers were divided by those by BioStage, and the ratio values represented CE. Results indicate that the CE of BioSampler was superior to that of Coriolis (P = 0.0001) and the PBS containing surfactant collected fungi better than that without surfactant (P < 0.0001), whereas antifoam agent showed no influence (P = 0.8). Moreover, fungal concentrations determined by BioSampler with surfactant‐added PBS were statistically indifferent from those by BioStage (P > 0.05) with a Spearman correlation coefficient of 0.81‐0.83 (P < 0.01). In addition to sampler and collection liquid, sampling location was also identified as a significant CE factor (P = 0.006), implying potential influences by fungal genera in the studied fields. Overall, BioSampler with surfactant‐supplemented PBS (eg, Triton X‐100) is recommended considering the great CE and compatibility with a variety of analytical assays.