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Development of a new bimodal imaging methodology: a combination of fluorescence microscopy and high‐resolution secondary ion mass spectrometry
Author(s) -
LAU K.H.,
CHRISTLIEB M.,
SCHRÖDER M.,
SHELDON H.,
HARRIS A.L.,
GROVENOR C.R.M.
Publication year - 2010
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2010.03380.x
Subject(s) - fluorescence microscope , secondary ion mass spectrometry , fluorescence , microscopy , mass spectrometry , fiducial marker , fluorescence lifetime imaging microscopy , resolution (logic) , analytical chemistry (journal) , chemistry , materials science , chromatography , optics , physics , artificial intelligence , computer science
Summary 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.