An in situ fracture device to image lipids in single cells using ToF-SIMS
2011 (English)In: Surface and Interface Analysis, ISSN 0142-2421, E-ISSN 1096-9918, Vol. 43, no 1-2, 257-260 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2011. Vol. 43, no 1-2, 257-260 p.
ToF-SIMS imaging, PC12 cells, freeze-fracture, phospholipids
IdentifiersURN: urn:nbn:se:uu:diva-228330DOI: 10.1002/sia.3542OAI: oai:DiVA.org:uu-228330DiVA: diva2:733730