期刊
JOURNAL OF MICROSCOPY
卷 240, 期 1, 页码 21-31出版社
WILEY-BLACKWELL
DOI: 10.1111/j.1365-2818.2010.03380.x
关键词
BrdU; cultured cells; fluorescence microscopy; NanoSIMS; secondary ion mass spectrometry (SIMS); subcellular localization; tetrathiomolybdate
类别
资金
- UK EPSRC [GR/T19797]
- Cancer Research UK [C5255/A8591]
- Engineering and Physical Sciences Research Council [GR/T19797/01] Funding Source: researchfish
P>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.
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