Fluorescence monitoring of antibiotic-induced bacterial damage using flow cytometry
✍ Scribed by M.T.E. Suller; D. Lloyd
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 214 KB
- Volume
- 35
- Category
- Article
- ISSN
- 0196-4763
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✦ Synopsis
Background: Conventional techniques used to assess bactericidal activities of antibodies are time-consuming; flow cytometry has been used as a rapid alternative. In this study, the membrane potential-sensitive fluorescent probes bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC 4 (3)) and Sytox Green, the redox dye cyano-2,3ditolyl tetrazolium chloride (CTC), and the Baclite viability test kit were used to assess the effects of ceftazidime, ampicillin, and vancomycin on clinical isolates of Pseudomonas aeruginosa, Escherichia coli, and Staphyloccocus aureus, respectively. Methods: Bacterial cultures were grown to early exponential phase, at which point the antibiotics were added at their breakpoint values, and incubation was allowed to continue. At timed intervals, samples were stained and flow cytometric analysis was performed on a Skatron Argus 100 arc-lamp based dual-parameter flow cytometer. Results: All the dyes successfully identified antibioticinduced damage in the three strains, although different fluorescence responses between the dyes were observed. DiBAC 4 (3) and Sytox Green overestimated numbers of