To cater to the diverse testing requirements of bioaerosol samplers in the market, and to advance the design, manufacturing, and practical application of these devices, this paper introduces two testing apparatuses — the fluorescence method and the counting method. These methods simulate microbial aerosols to assess the efficiency of microbial samplers. The composition, methodologies, and data processing steps of both testing devices are detailed. Subsequently, these methods were applied to evaluate the sampling efficiency of the domestically produced AGI-30 bioaerosol sampler and the Anderson six-stage microbial sampler under varying conditions. Results show that the AGI-30 sampler's efficiency increases with particle size, exceeding 90% for 5 μm particles, and decreases with higher flow rates. Sampling time also significantly impacts efficiency. The Anderson sampler exhibited high efficiency across eight particle sizes, approximately 99%, with a tendency to increase with particle size. The comparative analysis reveals that the fluorescence and counting methods, being more convenient than traditional microbial-culture methods, offer safe, reliable, and rapid results. The counting method, having fewer steps and no fluorescence quenching issues, is preferred for evaluating the efficiency of biological samplers.