Bioaerosol characterization by flow cytometry with fluorochrome

Traditional culture and microscopy methods for evaluation of bioaerosols are slow, tedious, and rather imprecise. In this study, the application of flow cytometry that was combined with a fluorescent technique (FCM/FL) was evaluated as a technique to quickly and accurately determine and quantify the total concentration and viability of bioaerosols. The optimal conditions of five fluorescent dyes [acridine orange (AO), SYTO-13, propidium iodide (PI), YOPRO-1, and 5-cyano-2,3-ditolytetrazolium chloride (CTC)] used in FCM/FL were determined for laboratory samples of bacterial aerosols (Escherichia coli, and endospores of Bacillus subtilis) and fungal aerosols (Candida famata and Penicillium citrinum spores). Based on the measured cell concentration, fluorescence intensity, and staining efficiency as indicators for dye performance evaluation, SYTO-13 was found to be the most suitable fluorescent dye for determining the total concentration of the bioaerosols, as well as YOPRO-1 was the most suitable for determining viability. Moreover, the established optimal FCM/FL with dyes was validated for characterizing microorganism profiles from both air and water samples from the aeration tank of hospital wastewater treatment plant. In conclusion, the FCM/FL successfully assessed the total concentration and viability for bacterial and fungal microorganisms in environmental field samples.