Microorganisms can be transmitted from infected to healthy people as an aerosol. Military bioaerosol detectors currently used by soldiers or first responders may potentially be utilized in health care settings as part of a strategy to prevent the spread of airborne infectious diseases. The goal of this study was to conduct initial laboratory and field validation of an inexpensive and unobtrusive TACBIO detector and compare its performance with that of an expensive bioaerosol detection instrument, the Ultraviolet Aerodynamic Particle Sizer (UV-APS). The laboratory validation test used three bacterial clusters (Bacillus thuringiensis [Bt], Bacillus anthracis Sterne [BaS], and Bacillus atrophaeus var. globigii [Bg]) generated at controlled rates by an ink jet aerosol generator (IJAG). The detection efficiency of the UV-APS was ≥ 99% for all particle generation rates and species. The TACBIO detector exhibited a slightly lower detection efficiency but was still able to detect > 88% of Bt and BaS and 62.7–81.7% of Bg. Field validation conducted with simultaneous UV-APS and TACBIO sampling in an occupied hospital clinic showed both instruments closely tracking each other in detecting fluorescent particles > 1.5 µm in diameter. During a 6 hour sampling period, fluorescent particle (> 1.5 µm) concentrations showed wide short term variation connected to nearby human activity while smaller nonfluorescent particles displayed more gradual changes. These results indicate the usefulness of an unobtrusive environmental aerosol sampler in health care settings, motivating future field characterization and validation studies.