Efficient quantification and characterization of bacterial outer membrane derived nano-particles with flow cytometric analysis

Andreas Wieser; Enno Storz; Gabriele Liegl; Annabell Peter; Michael Pritsch; Jonathan Shock; Sun Nyunt Wai; Sören Schubert

doi:10.1016/j.ijmm.2014.07.012

Efficient quantification and characterization of bacterial outer membrane derived nano-particles with flow cytometric analysis

Int J Med Microbiol. 2014 Nov;304(8):1032-7. doi: 10.1016/j.ijmm.2014.07.012. Epub 2014 Jul 27.

Authors

Andreas Wieser¹, Enno Storz², Gabriele Liegl², Annabell Peter², Michael Pritsch³, Jonathan Shock⁴, Sun Nyunt Wai⁵, Sören Schubert²

Affiliations

¹ Department of Bacteriology, Max von Pettenkofer Institute (LMU), Munich, Germany; Division of Infectious Diseases and Tropical Medicine, Medical Center of the University of Munich (LMU), Munich, Germany; German Center for Infection Reserach, Partner Site Munich, Munich, Germany; College of Public Health and Medical Science, Jimma University, Jimma, Ethiopia. Electronic address: wieser@mvp.uni-muenchen.de.
² Department of Bacteriology, Max von Pettenkofer Institute (LMU), Munich, Germany.
³ Department of Bacteriology, Max von Pettenkofer Institute (LMU), Munich, Germany; Division of Infectious Diseases and Tropical Medicine, Medical Center of the University of Munich (LMU), Munich, Germany; German Center for Infection Reserach, Partner Site Munich, Munich, Germany.
⁴ Laboratory for Quantum Gravity and Strings, Department of Mathematics and Applied Mathematics, University of Cape Town, Cape Town, South Africa.
⁵ Department of Molecular Biology and the Laboratory for Molecular Infection Medicine, Sweden (MIMS), Umeå University, Umeå SE-90187, Sweden.

PMID: 25139826
DOI: 10.1016/j.ijmm.2014.07.012

Abstract

There currently exists no efficient and easy method for size profiling and counting of membranous nano-scale particles, such as bacterial outer membrane vesicles (OMVs). We present here a cost-effective and fast method capable of profiling and counting small sample volumes of nano-scale membranous vesicles with standard laboratory equipment without the need for any washing steps. OMV populations of different bacterial species are compared and even subpopulations of OMVs can be identified after a simple labelling procedure. Counting is possible over three orders of magnitude without any changes to the protocol. Protein contaminations do not alter the described measurements.

Keywords: Escherichia coli; Flow cytometry; OMV; OMV enumeration; OMV quantification; Outer membrane vesicles.

Publication types

Evaluation Study

MeSH terms

Bacteria / chemistry*
Bacteriological Techniques / methods
Cell-Derived Microparticles / chemistry*
Flow Cytometry / methods*
Nanoparticles / analysis*