RESUMEN
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.
Asunto(s)
Diagnóstico por Imagen/métodos , Técnica del Anticuerpo Fluorescente/métodos , Microscopía Fluorescente/métodos , Espectrometría de Masa de Ion Secundario/métodos , Bromodesoxiuridina , Células Cultivadas , Células Endoteliales , Células HeLa , Humanos , Molibdeno , Cordón UmbilicalRESUMEN
This paper reviews the research in progress in Oxford on the chemical, spectroscopic and redox properties of bis(thiosemicarbazonato) complexes of zinc and copper in the context of the hypoxia selectivity of the copper(II) complex. Also, covered are synthetic strategies for modified complexes with a range of functional substituents and the in vitro and in vivo characteristics of two of these derivatives are described. Finally, the synthesis of some new bifunctional macrocyclic ligands is described and some of these give Cu(II) derivatives which cannot be reduced and are therefore resistant to reductive loss of copper in vivo.