Grant: Research Tools and Instruments

This application is for urgent replacement of a Multimode Microplate reader for fluorescence and luminescence . Our current instrument broke down 6 months ago and cannot be repaired. The equipment requested is essential to conduct our NSERC-funded research programs in Biochemistry, Cell Biology, Plant Functional Genomics and Environmental Toxicology and for student training in our laboratories. Our innovative research programs aim to: (1) understand cellular and molecular responses to different stresses (oxidative stress, heat shock, pesticides) in mammalian cells ; (2) develop efficient strategies to improve tolerance to oxidative stress, a major component of various abiotic and biotic stresses in plants ; (3) understand mechanisms by which environmental pollutants impair metabolic function and develop retinoid-based biomarkers for wildlife (e.g. bees) exposed to agricultural contamination ; and (4) provide molecular insights into the toxicology of trace elements in aquatic organisms using metallomic approaches . The equipment is requested by two senior (Averill-Bates, Houde), a mid-career (Boily) and a new investigator (Rosabal). These four programs all require analyses of fluorescence and luminescence in small sample volumes of mammalian cells; tissues and subcellular organelles from bees and aquatic organisms; and root tips and leaf tissues from plants. The Multimode Microplate Reader for fluorescence and luminescence comes with flexible wavelength selection for excitation and emission that allows enzyme kinetic analysis, endpoint measurements, and excitation/emission scans at any selected wavelength. It is equipped with top- and bottom-probe reading options for cell cultures, temperature control, and computer and software package for data processing. Luminescence, Time Resolved Fluorescence (TRF) and Fluorescence Resonance Energy Transfer (FRET) can also be detected. The instrument is essential for kinetic analysis of different proteases (caspases, calpains, lysosomal enzymes), acetylcholinesterase, phenoloxidase, and kinetics for generation of reactive oxygen species (superoxide, H 2 O 2 , NO ) and biomarkers of oxidative stress, using selective fluorescent substrates or probes. Of particular importance is the kinetic analysis of enzymatic reactions that requires rapid reading of samples at short time intervals to obtain initial reaction rates and kinetic parameters. Other applications include ELISA assays (albumin and insulin secretion) and sensitive detection of changes in proliferation, and lipid and protein oxidation, during genotoxic stress. Luminescence applications that are critical to our work include measurement of ATP, cAMP and cGMP. This equipment is indispensable for high quality, competitive research and graduate student training in the laboratories of four NSERC-funded investigators, at the Canadian and international levels.