Application of flow cytometry and cell sorting to the bacterial analysis of environmental aerosol samples.
Flow cytometry (FCM) combined with viability staining is a useful tool in discerning viable bacteria in environmental samples where traditional culture methods may fail. Contamination of aerosol samples with dust and other non-biological particles can interfere with accurate sample analysis and therefore there is a desire to exclude those particles from analysis. Particles were sorted according to their light scattering properties, cultured and isolates obtained. Isolates were cultured in suspension and reanalyzed by flow cytometry. The isolates were also analyzed and identified by DNA sequence analysis. Isolates with statistically similar light scattering properties shared common sequence identification. Isolates exhibited distinct light scattering profiles that roughly correlated with their originating gate, but often the peak of the profile was outside that gate.
United States Department of Agriculture, Agricultural Research Service, Western Regional Research Center, Foodborne Contaminants Research Unit, 800 Buchanan St., Albany, CA, USA. hernlemb@pw.usda.gov
Bradley J
BJ
Hernlem
Subbarao V
SV
Ravva
D000336:Aerosols; D000391:Air Microbiology; D000393:Air Pollutants / Q000302:isolation & purification*; D001419:Bacteria / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D002140:California; D015169:Colony Count, Microbial; D004784:Environmental Monitoring / Q000379:methods*; D005434:Flow Cytometry; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA
D000336:Aerosols; D000393:Air Pollutants; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S