Detachment of fungal spores from growing colonies results in human exposure. Thus far, the distribution of the binding forces of the spores in a fungal unit is unknown, so that precise prediction of the spores detachment is quite challenging. This investigation used centrifugal separation to measure the binding forces of the spores. Aspergillus niger (A. niger) colonies on a culture plate were placed in a centrifuge, the detached spores were counted, and this number was used to obtain the distribution of binding forces. Next, the air-blowing of an A. niger unit was modeled by computational fluid dynamics (CFD). A spore was judged to be detached if the air-imposed drag force was greater than the binding force. For model validation, the predicted spore detachment ratios were compared with the ratios measured in a wind tunnel test. The results revealed that the binding forces of the spores obeyed the log-normal distribution. The binding forces of the distal spores from colonies with a growth age of 66 h ranged from 0 nN to 4.0 nN and had a mean of 0.65 nN. The CFD modeling predicted the detachment ratios of the distal spores with good accuracy. •Binding forces of spores on the whole vesicle unit obey the log-normal distribution.•Relative deviation between the measured and predicted spore detachment ratios is <25 %.•Local air velocity surrounding a spore must be used to calculate the exerted drag force.