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Advanced air quality control requires real-time monitoring of particulate matter size and concentration, which can only be done using optical instruments. However, such techniques need regular calibration with reference samples. In this study, we suggest that puffball fungus ( Lycoperdon pyriforme ) spores can be utilized as a reference standard having a monodisperse size distribution. We compare the  Lycoperdon pyriforme  spores with the other commonly used reference samples, such as Al 2 O 3  powder and polystyrene latex (PSL) microspheres. Here we demonstrate that the puffball spores do not coagulate and, thus, maintain the same particle size in the aerosol state for at least 15 minutes, which is enough for instrument calibration. Moreover, the puffball mushrooms can be stored for several years and no agglomeration of the spores occurs. They are also much cheaper than other calibration samples and no additional devices are needed for aerosol generation since the fungal fruiting body acts as an atomizer itself. The aforementioned features make the fungal spores a highly promising substance for calibration and validation of particle size analyzers, which outperforms the existing, artificially produced particles for aerosol sampling. Furthermore, the  L. pyriforme  spores are convenient for basic research and development of new optical measurement techniques, taking into account their uniform particle size and absent coagulation in the aerosol.

Spores of puffball fungus Lycoperdon pyriforme as a reference standard of stable monodisperse aerosol for calibration of optical instruments