Microplate reader for fluorescence & luminescence

NSERC资助的四个生物化学和环境毒理学研究项目需要一台荧光酶标仪，其目的是了解：（1）细胞和分子对不同胁迫的反应（氧化应激，农药）;（2）重金属镉对成骨细胞增殖、迁移、分化、信号转导及凋亡机制的影响;和骨形成;（3）环境污染物（农药）损害动物组织和哺乳动物细胞代谢功能的机制;（4）抗氧化剂铜蓝蛋白在神经细胞和心肌细胞中的作用，以及研究其一氧化氮相关活性。这些项目都需要在细胞和组织的小样本体积中分析荧光和发光。2名高级调查员（D. Averill-Bates，J. Paquin）和2名初级研究人员（R.莫罗，M. Boily）。酶标仪具有灵活的激发和发射波长选择，允许动力学和终点测量，以及在任何选定波长下的激发和发射扫描。它配备了顶部和底部探头阅读选项、温度控制、计算机和数据处理软件包。还可以检测发光和荧光共振能量转移（FRET）。该设备对于使用选择性荧光底物参与凋亡信号传导和诱导细胞色素P450同工酶的不同蛋白酶（半胱天冬酶、钙蛋白酶、溶酶体酶）的动力学分析至关重要。其他应用包括ELISA测定（白蛋白和胰岛素分泌），以及在遗传毒性应激期间对增殖、钙、脂质和蛋白质氧化以及一氧化氮变化的灵敏检测。我们工作中感兴趣的发光应用包括测量细胞ATP、cAMP、cGMP以及神经元和骨分化的表型标记物的表达。这些设备对于在加拿大和国际一级的4名NSERC资助的研究人员的实验室中进行高质量，有竞争力的研究和研究生培训是不可或缺的。

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.