A simple and versatile methodology for high throughput toxicological assessment of chemical and environmental samples is presented. It uses panels of test organisms ranging from prokaryotic (E. coli, V. fischeri) and eukaryotic (Jurkat) cells to invertebrate (Artemia salina) and vertebrate (Danio rerio) organisms, to analyze alterations in their oxygen consumption by optical oxygen respirometry. All the assays are carried out in a convenient microtiter plate format using commercial reagents (phosphorescent oxygen probe, microplates) and detection on a standard fluorescent plate reader. Simple experimental set-up and mix-and-measure procedure allow parallel assessment of up to 96 samples (or assay points) in 2 h, easy generation of dose- and time-dependent responses, and EC(50) values. The methodology was demonstrated with several different classes of chemicals including heavy metal ions, PAHs, pesticides, their mixtures, and also validated with complex environmental samples such as wastewater from a wastewater treatment plant. It has been shown to provide high sensitivity, sample throughput and information content, flexibility and general robustness. It allows ranking and profiling of samples, compares favorably with alternative methods such as MicroTox and mortality tests with animal models, and is well suited for large-scale monitoring programs such as CWA and WFD.

We have optimized surface plasmon resonance (SPR) biosensor technology for a rapid, direct, and low-consumption label-free multianalyte screening of synthetic oligonucleotides (ONs) with modified internucleotide linkages potentially applicable in antisense therapy. Monitoring of the ONs hybridization is based on the formation of complex between the natural oligonucleotide probe immobilized on the sensor surface and the ON in solution in contact with the sensor surface. An immobilization chemistry utilizing the streptavidin-biotin interaction was employed to obtain desired ligand density and high hybridization efficiency. It was demonstrated that the sensor is capable of detecting complementary 23-mer ONs in concentrations as low as 0.1 nM with high specificity and reproducibility.

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Oxygen consumption is indicative of an organism's metabolic state, whereby alterations in respiration rate can result from the presence of different stimuli. Here, we develop a novel approach based on quenched fluorescence oxygen sensing and respirometry method for toxicity screening assays using the nematode Caenorhabditis elegans. Previously, C. elegans was established as a useful model in soil and aquatic toxicology studies. For existing toxicology screening approaches with C. elegans, however, the endpoint is lethality. In addition, the assay time frame for the existing approaches is considerably longer than that for the approach described here. We present a sensitive, robust, high-throughput platform using standard laboratory equipment for toxicological studies by measuring respiration rate in C. elegans animals using a phosphorescent probe.

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