Molecule specific imaging of freeze-fractured, frozen-hydrated model membrane systems using mass spectrometry

Donald M. Cannon, Michaeleen L. Pacholski, Nicholas Winograd, Andrew G. Ewing

Research output: Contribution to journalArticle

70 Scopus citations

Abstract

Imaging time-of-flight secondary ion mass spectrometry is used to chemically resolve the spatial distribution of lipids in submicrometer sections of phospholipid membranes. The results show that it is possible to unravel dynamical events such as chemical fluctuations associated with domain structure in cellular membranes. In this work, a liposome model system has been used to capture the stages of membrane fusion between two merging bilayer systems. Fracturing criteria for preserving chemical distributions are shown to be much more stringent than morphological electron cryomicroscopy studies. Images of membrane heterogeneity, induced via mixing various liposomes followed by fast freezing, demonstrate the necessary sample preparation groundwork to investigate complex, heterogeneous membrane domains. Clear delineation of membrane structure provides direct evidence that specific domains or 'rafts' can exist. Moreover, low concentrations of each phospholipid are distributed throughout newly fused liposomes despite the existence of distinct domains. In the liposome model, membrane structure ranges from specific domains to a fluid mosaic of the phospholipids during the fusion event. The availability of mass spectrometric imaging is proposed to facilitate the discovery of functional rafts or substructure in cell membranes before, during, and after events including cell division, exocytosis, endocytosis, intracellular transport, infection of membrane-bound viruses, and receptor clustering. This technology holds the promise to define the biology of cell membranes at the molecular level.

Original languageEnglish (US)
Pages (from-to)603-610
Number of pages8
JournalJournal of the American Chemical Society
Volume122
Issue number4
DOIs
StatePublished - Feb 2 2000

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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