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.
The original version of this chapter was revised. The erratum to this chapter is available at: DOI 10.1007/978-1-4939-6737-7_32
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-4939-6737-7_32
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Acknowledgments
We thank Jannik B. Larsen and Anncatrine L. Petersen for critical reading of the manuscript and valuable comments. Financial support for this work was kindly provided by the NanoMorph consortium—funded by the Danish Council for Technology and Innovation—and the Lundbeck Foundation Research Initiative on Brain Barriers and Drug Delivery.
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Kristensen, K., Henriksen, J.R., Andresen, T.L. (2017). Applying Fluorescence Correlation Spectroscopy to Investigate Peptide-Induced Membrane Disruption. In: Hansen, P. (eds) Antimicrobial Peptides. Methods in Molecular Biology, vol 1548. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6737-7_11
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DOI: https://doi.org/10.1007/978-1-4939-6737-7_11
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