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An in situ fracture device to image lipids in single cells using ToF-SIMS

Ingela Lanekoff ; Michael E. Kurczy (Institutionen för kemi- och bioteknik) ; Kelly L. Adams ; J Malm ; Roger Karlsson ; P Sjovall ; Andrew G Ewing
SURFACE AND INTERFACE ANALYSIS (0142-2421). Vol. 43 (2011), 1-2, p. 257-260.
[Artikel, refereegranskad vetenskaplig]

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.

Nyckelord: tof-sims imaging, pc12 cells, freeze-fracture, phospholipids

Denna post skapades 2012-08-17. Senast ändrad 2013-08-21.
CPL Pubid: 161982


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Institutionen för kemi (2001-2011)
Institutionen för kemi- och bioteknik (2005-2014)



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