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Article10.1007/978-1-62703-137-0_26

Secondary Ion Mass Spectrometry Imaging of Biological Membranes at High Spatial Resolution

Haley A. Klitzing,Peter Weber,Mary L. Kraft-2012-09-18-Methods in molecular biology
33

TL;DRAbstract

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.

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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.

Keywords

MembraneMass spectrometry imagingChemistrySecondary ion mass spectrometryMass spectrometryResolution (logic)SphingolipidCharacterization (materials science)

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