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In this paper, we present a new experimental methodology to combine mass spectrometry (NanoSIMS) with fluorescence microscopy to provide subcellular information on the location of small molecules in cultured cells. We demonstrate this by comparing the distribution of 5-bromo-2-deoxyuridine in the same cells given by both NanoSIMS analysis and by fluorescence immunohistochemistry. Fiducial markers in the substrates ensured that the images formed by SIMS mapping of bromine ions could be co-registered exactly with images from fluorescence microscopy. The NanoSIMS was shown to faithfully reproduce the information from fluorescence microscopy, but at a much higher spatial resolution. We then show preliminary SIMS images on the distribution of ATN-224, a therapeutic copper chelator for which there is no fluorescent marker, co-registered with conventional Lysotracker and Hoechst stains on the same cells.

Original publication

DOI

10.1111/j.1365-2818.2010.03380.x

Type

Journal article

Journal

J Microsc

Publication Date

10/2010

Volume

240

Pages

21 - 31

Keywords

Bromodeoxyuridine, Cells, Cultured, Diagnostic Imaging, Endothelial Cells, Fluorescent Antibody Technique, HeLa Cells, Humans, Microscopy, Fluorescence, Molybdenum, Spectrometry, Mass, Secondary Ion, Umbilical Cord