Identification of coexistence of biological and non-biological aerosol particles with DAPI (4′,6-diamidino-2-phenylindole) stain

Abstract

The fluorescent dye 4′,6-diamidino-2-phenylindole (DAPI) has been widely used to stain microorganisms in various environment media. We applied DAPI fluorescence enumeration to airborne microorganisms and found that non-biological particles, including organic compounds, minerals, and soot, were also visible upon exposure to UV excitation under fluorescence microscope. Using laboratory-prepared biological particles as the control, we investigated the feasibility of identifying both biological and non-biological particles in the same sample with DAPI staining. We prepared biological (bacterial, fungi, and plant detritus) and non-biological (biochar, soot, mineral, metal, fly ash, salt) particles in the laboratory and enumerated the particles and their mixture with DAPI. We found that mineral particles were transparent, and biochar, soot, metals and fly ash particles were black under a filter set at excitation 350/50 nm and emission 460/50 nm bandpass (DAPI-BP), while biological particles were blue, as expected. Particles of the water-soluble salts NaCl and (NH4)2SO4 were yellow under a filter set at excitation 340–380 nm and emission 425 nm long pass (DAPI-LP). Case studies with samples of dustfall, atmospheric aerosols and surface soils could allow for the quantification of the relative number of different types of particles and particles with organic matter or salt coating as well. Fluorescence enumeration with DAPI stain is thus able to identify the co-existence of biological and non-biological particles in the air, at least to the extent of those examined in this study.