期刊
SURFACE AND INTERFACE ANALYSIS
卷 43, 期 1-2, 页码 257-260出版社
WILEY
DOI: 10.1002/sia.3542
关键词
ToF-SIMS imaging; PC12 cells; freeze-fracture; phospholipids
资金
- National Institutes of Health
- Swedish Research Council
- European Union
Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) imaging yields molecule-specific images showing the spatial distribution of specific lipids with submicron resolution, making it a potentially powerful method for studying a variety of biological questions in single cells. In order to make possible the analysis of hydrated cells in vacuum, we have designed a device for in situ freeze-fracture of cell samples. PC12 cells are frozen between two silicon shards in a sandwich geometry, and the fracture is conducted under vacuum inside the analysis chamber of the instrument. After freeze-fracture, ToF-SIMS analysis was carried out using a Bi cluster ion source, providing high-resolution images of single cells. We also show that when combining freeze-fracture and cluster SIMS imaging, low abundant biologically important lipids, specifically PE, PC and non-SM PC, can be imaged in single cells. Finally, K+ ion localization was used as a diagnostic to identify fracture planes through the cell for these fractured samples. Copyright (C) 2010 John Wiley & Sons, Ltd.
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