Secondary ion mass spectrometry imaging of biological membranes at high spatial resolution

Haley A. Klitzing, Peter K. Weber, Mary L. Kraft

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Characterization of the distributions of specific proteins and lipids within cellular membranes is currently a major challenge. Advances in secondary ion mass spectrometry (SIMS) now enable the distributions of isotopically labeled lipids within cellular or model membranes to be imaged with chemical specificity and high (≥50 nm) lateral resolution. Here, methods to image the distributions of sphingolipids within the membranes of intact cells with a Cameca NanoSIMS are described. For NanoSIMS detection, the incorporation of distinct stable isotopes into the lipid species of interest is essential. Metabolic labeling, cell preservation, imaging conditions, and data analysis are critical factors. The methods and principles described here can be extended to studying other membrane lipids or cholesterol.

Original languageEnglish (US)
Title of host publicationNanoimaging
Subtitle of host publicationMethods and Protocols
EditorsAlioscka Sousa, Michael Kruhlak
Pages483-501
Number of pages19
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume950
ISSN (Print)1064-3745

Keywords

  • Chemical imaging
  • Lipid organization
  • Metabolic labeling
  • NanoSIMS
  • Secondary ion mass spectrometry

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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