Protocol

Mass Spectrometry Imaging

Volume 656 of the series Methods in Molecular Biology pp 85-97

Date:

Lipid Detection, Identification, and Imaging Single Cells with SIMS

  • Michael L. HeienAffiliated withDepartment of Chemistry and Biochemistry, The University of Arizona
  • , Paul D. PiehowskiAffiliated withDepartment of Chemistry, The Pennsylvania State University
  • , Nicholas WinogradAffiliated withDepartment of Chemistry, The Pennsylvania State University
  • , Andrew G. EwingAffiliated withDepartment of Chemistry, The Pennsylvania State UniversityDepartment of Chemistry, Göteborg University

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Abstract

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) can be utilized to map the distribution of various molecules on a surface with submicrometer resolution. Many of its biological applications have been in the study of membrane lipids, such as phospholipids and cholesterol. For these studies, the effectiveness of chemical mapping is limited by low signal intensity from various biomolecules. Because of the high-energy nature of the SIMS ionization process, many molecules are identified by detection of characteristic fragments. Cluster ion sources are able to increase ionization, leading to increased information collected from a surface. In this chapter, we highlight the utility of SIMS to image lipids at single cells. Particularly, we will describe sample preparation, data collection, and the analysis of lipids for two systems; rat oligodendrocytes and Tetrahymena thermophila. SIMS spectra yield information regarding lipid identity and concentration across cell surface.

Key words

Time-of-flight (ToF) secondary ion mass spectrometry (SIMS) freeze-fracture freeze-etch imaging mass spectrometry lipids single cell