BackgroundHumans emit approximately 30 million microbial cells per hour into their immediate vicinity. However, sampling of aerosolized microbial taxa (aerobiome) remains largely uncharacterized due to the complexity and limitations of sampling techniques, which are highly susceptible to low biomass and rapid sample degradation. Recently, there has been an interest in developing technology that collects naturally occurring water from the atmosphere, even within the built environment. Here, we analyze the feasibility of indoor aerosol condensation collection as a method to capture and analyze the aerobiome.MethodsAerosols were collected via condensation or active impingement in a laboratory setting over the course of 8 h. Microbial DNA was extracted from collected samples and sequenced (16S rRNA) to analyze microbial diversity and community composition. Dimensional reduction and multivariate statistics were employed to identify significant (p< 0.05) differences in relative abundances of specific microbial taxa observed between the two sampling platforms.ResultsAerosol condensation capture is highly efficient with a yield greater than 95% when compared to expected values. Compared to air impingement, aerosol condensation showed no significant difference (ANOVA,p> 0.05) in microbial diversity. Among identified taxa, Streptophyta and Pseudomonadales comprised approximately 70% of the microbial community composition.ConclusionThe results suggest that condensation of atmospheric humidity is a suitable method for the capture of airborne microbial taxa reflected by microbial community similarity between devices. Future investigation of aerosol condensation may provide insight into the efficacy and viability of this new tool to investigate airborne microorganisms.ImportanceOn average, humans shed approximately 30 million microbial cells each hour into their immediate environment making humans the primary contributor to shaping the microbiome found within the built environment. In addition, recent events have highlighted the importance of understanding how microorganisms within the built environment are aerosolized and dispersed, but more importantly, the lack in development of technology that is capable of actively sampling the ever-changing aerosolized microbiome, i.e., aerobiome. This research highlights the capability of sampling the aerobiome by taking advantage of naturally occurring atmospheric humidity. Our novel approach reproduces the biological content in the atmosphere and can provide insight into the environmental microbiology of indoor spaces.Video AbstractSupplementary InformationThe online version contains supplementary material available at 10.1186/s40168-023-01555-5.