ArticleNature protocols2018
Spectrofluorimetric quantification of antibiotic drug concentration in bacterial cells for the characterization of translocation across bacterial membranes.
Article in Nature protocols, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
29 citing papers in PubMed, 54 citations in OpenAlex.
- NV716 acts as an envelope-active adjuvant that enhances antibiotic accumulation in Pseudomonas aeruginosa.npj antimicrobials and resistance · 2026Article
- Click-based determination of accumulation of molecules in Escherichia coli.Nature communications · 2026Article
- Antechodynamics and Antechokinetics: Dynamics and Kinetics of Antibiotic Resistance Biomolecules.Biomolecules · 2025Review
- Unlocking the Gates: A Novel Diagnostic Molecule for Quantifying Efflux Levels in Gram-Positive Bacteria.Advanced healthcare materials · 2025Article
- Article
- Advances in methods and concepts provide new insight into antibiotic fluxes across the bacterial membrane.Communications biology · 2024Article
- Role of the Bacterial Amyloid-like Hfq in Fluoroquinolone Fluxes.Microorganisms · 2023Article
- A framework for dissecting affinities of multidrug efflux transporter AcrB to fluoroquinolones.Communications biology · 2022Article
- Cephalosporin translocation across enterobacterial OmpF and OmpC channels, a filter across the outer membrane.Communications biology · 2022Article
- Article
- An Outer Membrane Vesicle-Based Permeation Assay (OMPA) for Assessing Bacterial Bioavailability.Advanced healthcare materials · 2022Article
- An LC-MS/MS assay and complementary web-based tool to quantify and predict compound accumulation in E. coli.Nature protocols · 2021Review
- Clinical Status of Efflux Resistance Mechanisms in Gram-Negative Bacteria.Antibiotics (Basel, Switzerland) · 2021Review
- Tolerance engineering in Deinococcus geothermalis by heterologous efflux pumps.Scientific reports · 2021Article
- Design and Clinical Application of an Integrated Microfluidic Device for Circulating Tumor Cells Isolation and Single-Cell Analysis.Micromachines · 2021Article
- Exploring the Chemical Space of Benzothiazole-Based DNA Gyrase B Inhibitors.ACS medicinal chemistry letters · 2020Article
- Defining new chemical space for drug penetration into Gram-negative bacteria.Nature chemical biology · 2020Review
- Fluorescent macrolide probes - synthesis and use in evaluation of bacterial resistance.RSC chemical biology · 2020Article
- Single-cell microfluidics facilitates the rapid quantification of antibiotic accumulation in Gram-negative bacteria.Lab on a chip · 2020Article
- Synchrotron multimodal imaging in a whole cell reveals lipid droplet core organization.Journal of synchrotron radiation · 2020Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The efficacy of antibacterial molecules depends on their capacity to reach inhibitory concentrations in the vicinity of their target. This is particularly challenging for drugs directed against Gram-negative bacteria, which have a complex envelope comprising two membranes and efflux pumps. Precise determination of the bacterial drug content is an essential prerequisite for drug development. Here we describe three approaches that have been developed in our laboratories to quantify drugs accumulated in intact cells by spectrofluorimetry, microspectrofluorimetry, and kinetics microspectrofluorimetry (KMSF). These different procedures provide complementary results that highlight the contribution of membrane-associated mechanisms, including influx through the outer membrane (OM) and efflux, and the importance of the physicochemical properties of the transported drugs for the intracellular concentration of a given antibiotic in a given bacterial population. The three key stages of this protocol are preparation of the bacterial strains in the presence of the antibiotic; preparation of the whole-cell lysates (WCLs) and fluorescence readings; and data analysis, including normalization and quantitation of the intracellular antibiotic fluorescence relative to the internal standard and the antibiotic standard curve, respectively. Fluorimetry is limited to naturally fluorescent or labeled compounds, but in contrast to existing alternative methods such as mass spectrometry, it uniquely allows single-cell analysis. From culture growth to data analysis, the protocol described here takes 5 d.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.