Bioaerosol characterization by flow cytometry with fluorochrome.

Area of Science:

• Environmental microbiology
• Analytical chemistry
• Biotechnology

Background:

• Traditional bioaerosol evaluation methods (culture and microscopy) are time-consuming and lack precision.
• Accurate quantification and viability assessment of bioaerosols are crucial for environmental and health monitoring.

Purpose of the Study:

• To evaluate flow cytometry combined with fluorescence (FCM/FL) as a rapid and accurate technique for bioaerosol analysis.
• To determine optimal fluorescent dyes for quantifying total bioaerosol concentration and viability.
• To validate the FCM/FL method for environmental samples.

Main Methods:

• Optimized conditions for five fluorescent dyes (AO, SYTO-13, PI, YOPRO-1, CTC) using laboratory bacterial (E. coli, Bacillus subtilis) and fungal (Candida famata, Penicillium citrinum) aerosols.
• Evaluated dye performance based on cell concentration, fluorescence intensity, and staining efficiency.
• Validated the optimized FCM/FL method on air and water samples from a hospital wastewater treatment plant aeration tank.

Main Results:

• SYTO-13 was identified as the most suitable dye for determining total bioaerosol concentration.
• YOPRO-1 proved most effective for assessing bioaerosol viability.
• The optimized FCM/FL method successfully characterized microorganism profiles in real-world environmental samples.

Conclusions:

• FCM/FL provides a significant advancement over traditional methods for bioaerosol assessment.
• The validated FCM/FL technique accurately quantifies both total concentration and viability of airborne and waterborne bacteria and fungi.
• This method has strong potential for environmental monitoring and public health applications.