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An in situ fracture device to image lipids in single cells using ToF‐SIMS
Author(s) -
Lanekoff Ingela,
Kurczy Michael E.,
Adams Kelly L.,
Malm Jakob,
Karlsson Roger,
Sjövall Peter,
Ewing Andrew G.
Publication year - 2011
Publication title -
surface and interface analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.52
H-Index - 90
eISSN - 1096-9918
pISSN - 0142-2421
DOI - 10.1002/sia.3542
Subject(s) - in situ , fracture (geology) , secondary ion mass spectrometry , resolution (logic) , cluster (spacecraft) , ion , high resolution , chemistry , mass spectrometry , analytical chemistry (journal) , materials science , composite material , geology , computer science , chromatography , remote sensing , organic chemistry , artificial intelligence , programming language
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 © 2010 John Wiley & Sons, Ltd.