Single Cell Flow Cytometry Assay for Peptide Uptake by Bacteria.
Monica Benincasa, Quentin Barrière, Giulia Runti, Olivier Pierre, Mick Bourge, Marco Scocchi, Peter Mergaert
Author Information
Monica Benincasa: Department of Life Sciences, University of Trieste, Trieste, Italy.
Quentin Barrière: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette cedex, France.
Giulia Runti: Department of Life Sciences, University of Trieste, Trieste, Italy.
Olivier Pierre: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette cedex, France.
Mick Bourge: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette cedex, France.
Marco Scocchi: Department of Life Sciences, University of Trieste, Trieste, Italy.
Peter Mergaert: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette cedex, France.
Antimicrobial peptides (AMPs) can target the bacterial envelope or alternatively have intracellular targets. The latter requires uptake of the peptide by the bacterial cells. The bacterial internalization of an AMP can be evaluated by a fluorescence-based method that couples the use of the fluorescently labelled AMP to the fluorescence quencher trypan blue. Trypan blue is excluded from the interior of intact cells and the fluorescence of the extracellular peptide or of the peptide bound on the bacterial surface can be quenched by it, while the fluorescence of the internalized peptide is not affected. The uptake of the peptide by the bacteria is determined by measuring the fluorescence in individual cells by flow cytometry.
