A Single‐Molecule Liposome Assay for Membrane Permeabilization
Abstract
Abstract Cell membrane disruption is associated with numerous diseases and underlies the activity of various antimicrobial agents. The rapid screening of compounds capable of disrupting or permeabilizing biological membranes is essential to the search for new therapeutic drugs. Here, we present a single‐molecule confocal microscopy assay integrated with fast‐flow microfluidics to study membrane permeabilization in large unilamellar vesicles (LUVs) containing as few as seven dye molecules. This assay eliminates the need for liposome immobilization and achieves detection rates in the range of 1000 vesicles per minute, offering unparalleled sensitivity and detection limits as low as 135 pM, corresponding to just eight permeabilizing molecules per vesicle for active compounds such as ionomycin. It provides a robust platform for investigating membrane‐disrupting agents, including those with antimicrobial properties or implicated in neurodegenerative diseases.
Article Details
Authors (10)
Krzysztof M. Bąk
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK
Daniel C. Edwards
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK
Dylan George
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK
Bhanu Singh
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK
Ryan Ferguson
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK
Tianxiao Zhao
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK
Kristin Piché
StressMarq Biosciences Inc., 118-1537 Hillside Avenue, Victoria BC V8T 2C1, British Columbia
Ariel Louwrier
Scott L. Cockroft
EaStCHEM School of Chemistry, University of Edinburgh, Joseph Black Building, David Brewster Rd, Edinburgh, EH9 3FJ, United Kingdom
Mathew H. Horrocks
EaStCHEM School of Chemistry University of Edinburgh Joseph Black Building, David Brewster Rd Edinburgh EH9 3FJ UK