EuroTracker Dyes: Synthesis and Application in Functional Cell Imaging

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The visualisation of certain living cells or microbes by optical microscopy requires the introduction of bright, luminescent dyes or stains, that can light up selected parts of the cell effectively.We will develop bright complexes of the rare earth element, europium, (EuroTracker dyes) as optical probes that emit light efficiently and report information back to the observer about their chemical or biological environment. These probes have been designed to allow them to be excited by the light sources that are available in microscopes used by physical scientists, biochemists and cell biologists, to study the interior composition of cells or related small compartments. By examining series of closely related probes, we shall define systems that locate to a particular compartment of a cell, allowing it to be visualised selectively. The optical probes that are currently used are based either on organic compounds, on large man-made proteins or on so-called quantum dots; each of these probes suffer from problems of low chemical stability, a tendency to fade and, in certain cases, they are rather toxic to the living cell. Thus we aim to change the thinking of the microscopy community and to pave the way for such emissive metal complexes to be used as stains and probes in biology. These metal complex probes emit light over longer timescales than conventional probes, and so their is more time to collect the emitted light after pulsed excitation and only short exposure times to the light that is used to excite them. The long emission lifetime avoids problems in data acquisition that are associated with light scattering and background autofluorescence. In parallel, we shall develop instrumentation methods that allow the spectral signature of these probes to be recorded quickly using fast imaging methods. We shall work with microscope manufacturers and with optics specialists to this end. This will allow changes in chemical composition within a cell to be tracked on a 10 second timescale. As a first step, we shall develop probes that can report on pH change and on the amount of sodium, magnesium and zinc within a specific cell compartment, allowing changes in the amount of these important metal ions to be tracked in living cells in real time. By adopting a modular approach to probe design, different metal ions can be examined subsequently, by introducing the appropriate metal-binding moiety into the probe structure.