Grant: Research Tools and Instruments - Category 1 (<$150,000)

A Microplate Reader for Fluoresence and Luminescence is requested for four NSERC-funded research programs in Biochemistry and Environmental Toxicology, which aim to understand: (1) the cellular and molecular responses to different stresses (oxidative stress, pesticides); (2) the impact of heavy metal cadmium on mechanisms involved in the proliferation, migration, differentiation and signal transduction, as well as apoptosis of osteoblastic cells, and bone formation; (3) mechanisms by which environmental pollutants (agricultural pesticides) impair metabolic function in animals tissues and mammalian cells; and (4) the action of the antioxidant ceruloplasmin in neuronal cells and cardiomyocytes, as well as investigation of its nitric oxide-related activities. These projects all require analyses of fluorescence and luminescence in small sample volumes of cells and tissues. The equipment is requested by 2 senior investigators (D. Averill-Bates, J. Paquin) and 2 junior researchers (R. Moreau, M. Boily). The Microplate Reader comes with flexible wavelength selection for excitation and emission that allows kinetics and endpoint measurements, and excitation and emission scans at any selected wavelength. It is equipped with top- and bottom-probe reading options, temperature control, computer and software package for data processing. Luminescence and Fluorescence Resonance Energy Transfer (FRET) can also be detected. This equipment is essential for kinetics analysis of different proteases (caspases, calpains, lysosomal enzymes) involved in apoptotic signalling, and induction of cytochrome P450 isoenzymes, using selective fluorescent substrates. Other applications include ELISA assays (albumin and insulin secretion), and sensitive detection of changes in proliferation, calcium, lipid and protein oxidation and nitric oxide, during genotoxic stress. Luminescence applications of interest to our work include measurement of cellular ATP, cAMP, cGMP, and expression of phenotypic markers of neuronal and bone differentiation. This equipment is indispensable for high quality, competitive research and graduate student training in laboratories of 4 NSERC-funded investigators, at the Canadian and international levels.