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In this brief review, we present recent results in the development of fluorescence-based assays for the detection of compounds with cytotoxic, anticancer and anti-microbial properties. As other reviews have explored various aspects related to these topics, this review will focus on the use of the Green Fluorescent Protein (GFP) for the detection of potentially toxic and/or therapeutic compounds. Since high-throughput screening of chemical compounds can be both expensive and laborious, development of low cost and efficient cell-based assays to determine biological activity should greatly enhance the early screening process. In our recent studies, we have developed a couple of GFP-based assays for the rapid screening of compounds with cytotoxic and bacteriocidal properties. As will be described in more detail in subsequent sections, a new 96-well assay has recently been developed that allows for the simultaneous screening of test compounds on gram positive and negative bacteria as well as mammalian (human cancer) cells. Our results demonstrate that both mammalian cells and bacteria can be analyzed in tandem to rapidly determine which compounds are specifically toxic to one of these cell types. The parallel screening of both eukaryotic and prokaryotic cells was found to be feasible, reproducible, and cost effective.

Following the considerable impact of the application of convenient ultrafast lasers to multiphoton microscopy on biomedical imaging, it seems to us that FLIM and MDFI continue the trend in which advances in instrumentation will facilitate new discoveries — and modes of discovery — in biology and medicine. We hope we have shown the reader that fluorescence lifetime can provide intrinsic molecular contrast in unstained tissue and that the prospects for in vivo application are exciting. We believe that the capability to excite fluorophores at almost any excitation wavelength and the opportunities to extract more information from fluorescence signals by resolving with respect to lifetime, excitation and emission spectrum and also polarisation, will have a major impact on the ability to identify and exploit intrinsic contrast and on investigations of molecular biology. There the combination of new fluorescence probe technology, including genetically-expressed labels and nano-engineered devices, with new modes of interrogation and analysis, will continue to fuel the astounding advances in this field. There is a real prospect that our ability to ask and test biological questions will cease to be limited by the availability of suitable instrumentation. Rather it is likely to be limited by our ability to analyse and comprehend the (rapidly increasing volume of) data that we collect.