title: Applying Fluorescence Correlation Spectroscopy to Investigate Peptide-Induced Membrane Disruption.
abstract: There is considerable interest in understanding the interactions of antimicrobial peptides with phospholipid membranes. Fluorescence correlation spectroscopy (FCS) is a powerful experimental technique that can be used to gain insight into these interactions. Specifically, FCS can be used to quantify leakage of fluorescent molecules of different sizes from large unilamellar lipid vesicles, thereby providing a tool for estimating the size of peptide-induced membrane disruptions. If fluorescently labeled lipids are incorporated into the membranes of the vesicles, FCS can also be used to obtain information about whether leakage occurs due to localized membrane perturbations or global membrane destabilization. Here, we outline a detailed step-by-step protocol on how to optimally implement an FCS-based leakage assay. To make the protocol easily accessible to other researchers, it has been supplemented with a number of practical tips and tricks.
authors.affiliation: Department of Micro- and Nanotechnology, DTU Nanotech, Technical University of Denmark, Kgs. Lyngby, Denmark.|Center for Nanomedicine and Theranostics, Technical University of Denmark, Kgs. Lyngby, Denmark.
authors.forename: Kasper
authors.initials: K
authors.lastname: Kristensen
authors.affiliation: Center for Nanomedicine and Theranostics, Technical University of Denmark, Kgs. Lyngby, Denmark.|Department of Chemistry, DTU Chemistry, Technical University of Denmark, Kgs. Lyngby, Denmark.
authors.forename: Jonas R
authors.initials: JR
authors.lastname: Henriksen
authors.affiliation: Department of Micro- and Nanotechnology, DTU Nanotech, Technical University of Denmark, Kgs. Lyngby, Denmark. tlan@nanotech.dtu.dk.|Center for Nanomedicine and Theranostics, Technical University of Denmark, Kgs. Lyngby, Denmark. tlan@nanotech.dtu.dk.
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Andresen
mesh_terms: D000465:Algorithms; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D008051:Lipid Bilayers; D008563:Membrane Lipids; D010455:Peptides; D010743:Phospholipids
keywords: Antimicrobial peptides; Fluorescence correlation spectroscopy (FCS); Lipid vesicles; Membrane disruption; Membrane-active peptides; Peptide-lipid membrane interactions; Pore formation; Vesicle leakage assay