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Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth
Published on: November 24, 2017
Cytoplasmic Accumulation and Permeability of Antibiotics in Gram Positive and Gram Negative Bacteria Visualized in
Scott O Squire1,2, Sepehr Sebghati1,2, Ming C Hammond1,2
1Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, United States.
Abstract:
In this study, we describe the first real-time live cell assay for compound accumulation and permeability in both Gram positive and Gram negative bacteria. The assay utilizes a novel fluorogenic tagging strategy that permits direct visualization of compound accumulation dynamics in the cytoplasm of live cells, unobscured by washing or other processing steps. Quantitative differences could be reproducibly measured by flow cytometry at compound concentrations below the limit of detection for MS-based approaches. We establish the fluorogenic assay in E. coli and B. subtilis and compare the intracellular accumulation of two antibiotics, ciprofloxacin and ampicillin, with related pharmacophores in these bacteria.
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