ToF-SIMS imaging and depth profiling of HeLa cells treated with bromodeoxyuridine

Citation

Brison, J.; Benoit, D. S. W.; Muramoto, S.; Robinson, M.; Stayton, P. S.; & Castner, D. G. (2011). ToF-SIMS imaging and depth profiling of HeLa cells treated with bromodeoxyuridine. Surface and Interface Analysis, 43(1-2), 354-357.

Abstract

Time-of-flight (ToF) SIMS 2D images and molecular depth profiles of human HeLa cells treated with bromodeoxyuridine (BrdU) were acquired in the dual beammode (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 degrees. and rastered over a 500 x 500 mu m(2) area, similar to 1 nm of biological material was sputtered every second. Our results show that HeLa cells were completely removed after etching with 1.3 x 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 degrees. incidence. Copyright (C) 2010 John Wiley & Sons, Ltd.

Keyword(s)

brdu
bromodeoxyuridine
c(60)
challenges
cultures
depth profiling
dual beam
elements
hela cells
imaging
ion mass-spectrometry
molecules
resolution
sample preparation
single cells
tissues
tof-sims

Notes

725TR
Times Cited:16
Cited References Count:18

Reference Type

Journal Article

Secondary Title

Surface and Interface Analysis

Author(s)

Brison, J.
Benoit, D. S. W.
Muramoto, S.
Robinson, M.
Stayton, P. S.
Castner, D. G.

Year Published

2011

Volume Number

43

Issue Number

1-2

Pages

354-357

DOI

Doi 10.1002/Sia.3415