Secondary Ion MS Imaging To Relatively Quantify Cholesterol in the Membranes of Individual Cells from Differentially Treated Populations

Sara G. Ostrowski, Michael E. Kurczy, Thomas P. Roddy, Nicholas Winograd, and Andrew G. Ewing*
Department of Chemistry, 104 Chemistry Research Building, Pennsylvania State University, University Park, Pennsylvania 16802
Anal. Chem., 2007, 79 (10), pp 3554–3560
DOI: 10.1021/ac061825f
Publication Date (Web): April 12, 2007
Copyright © 2007 American Chemical Society
*

 Corresponding author. E-mail:  age@psu.edu.

Abstract

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) is a well-established bioanalytical method for directly imaging the chemical distribution across single cells. Here we report a protocol for the use of SIMS imaging to comparatively quantify the relative difference in cholesterol level between the plasma membranes of two cells. It should be possible to apply this procedure to the study of other selected lipids. This development enables direct comparison of the chemical effects of different drug treatments and incubation conditions in the plasma membrane at the single-cell level. Relative, quantitative TOF-SIMS imaging has been used here to compare macrophage cells treated to contain elevated levels of cholesterol with respect to control cells. In situ fluorescence microscopy with two different membrane dyes was used to discriminate morphologically similar but differentially treated cells prior to SIMS analysis. SIMS images of fluorescently identified cells reveal that the two populations of cells have distinct outer leaflet membrane compositions with the membranes of the cholesterol-treated macrophages containing more than twice the amount of cholesterol of control macrophages. Relative quantification with SIMS to compare the chemical composition of single cells can provide valuable information about normal biological functions, causative agents of diseases, and possible therapies for diseases.

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History

  • Published In Issue May 15, 2007
  • Received for review September 27, 2006. Accepted March 13, 2007.

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