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ToF‐SIMS imaging and depth profiling of HeLa cells treated with bromodeoxyuridine
Author(s) -
Brison Jeremy,
Benoit Danielle S. W.,
Muramoto Shin,
Robinson Michael,
Stayton Patrick S.,
Castner David 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.3415
Subject(s) - hela , chemistry , analytical chemistry (journal) , bromodeoxyuridine , etching (microfabrication) , ion beam , materials science , ion , cell , nanotechnology , chromatography , cell growth , biochemistry , organic chemistry , layer (electronics)
Time‐of‐flight (ToF) SIMS 2D images and molecular depth profiles of human HeLa cells treated with bromodeoxyuridine (BrdU) were acquired in the dual beam mode (Bi 3 + analysis beam, C 60 + etching beam). Several preparation protocols were investigated and were compared to a simple wash‐and‐dry method. The feasibility of using C 60 to clean the samples prior to imaging with Bi was also investigated quantitatively by calibrating full depth profiles of the cells using atomic force microscopy. BrdU was used as a marker for the cell nucleus, facilitating identification and localization of subcellular features during depth profiling. Results show that C 60 can be used to remove the surface contamination and to access different layers within the cells for 2D imaging. For a 1 nA, 10 keV C 60 + beam incident at 45° and rastered over a 500 × 500 µm 2 area, ∼1 nm of biological material was sputtered every second. Our results show that HeLa cells were completely removed after etching with 1.3 × 10 15 C 60 + ions per cm 2 , giving an average etching rate of 3.9 nm for every 10 13 C 60 per cm 2 at 10 keV and 45° incidence. Copyright © 2010 John Wiley & Sons, Ltd.