A Phosphorimager for Research in Plant Biology

植物生物学系和农业生物技术中心的一组研究人员要求提供一台多用户磷光成像仪。该仪器通常被称为快速、准确、无胶片放射自显影系统。它有助于快速和灵敏地定量凝胶或印迹上的放射性，而无需闪烁计数或放射自显影。高灵敏度的存储荧光屏捕捉实验样品的图像比胶片快十倍。磷光成像仪的宽线性动态范围允许以胶片根本无法实现的方式定量检测强信号和弱信号。用微机对数字图像进行编辑和分析。我们建议使用这种强大的仪器在广泛的应用，包括定量分析的DNA-蛋白质相互作用的凝胶阻滞，测量转录水平的RNA凝胶印迹或RNase保护试验，和动力学测量酶活性和蛋白质磷酸化。磷光成像仪的获得将使我们能够获得定量信息和研究问题的答案，否则使用传统的X射线胶片和放射自显影是不可能获得的。七名调查人员中有六名目前得到了联邦机构的竞争性赠款的支持。在所有情况下，所提出的实验需要分析的磷成像仪是必不可少的长期目标，了解植物发病机制，信号转导，膜运输和有丝分裂的分子基础。

A multi-user phosphorimager is requested by a group of investigators in the Department of Plant Biology and the Center for Agricultural Biotechnology. This instrument is often referred to as a fast, accurate, filmless autoradiography system. It facilitates the rapid and sensitive quantitation of radioactivity on gels or blots without scintillation counting or autoradiography. Highly sensitive storage phosphor screens capture the image of the experimental sample ten times faster than film. The broad linear dynamic range of the phosphoimager permits quantitative detection of both strong and weak signals in a way that film simply cannot. Digital images are compiled and analyzed with a microcomputer. We propose to use this powerful instrumentation in a wide range of applications, including the quantitative analyses of DNA-protein interactions by gel retardation, measurement of transcript levels by RNA gel blots or RNase protection assays, and kinetic measurements of enzyme activity and protein phosphorylation. The acquisition of a phosphorimager will permit us to obtain quantitative information and answers to research questions that would otherwise be impossible to obtain using conventional X-ray film and autoradiography. Six of the seven investigators are currently supported by competitive grants from federal agencies. In all cases, the proposed experiments requiring analyses by the phosphorimager are essential to the long-range goals of understanding the molecular basis for plant pathogenesis, signal transduction, membrane transport and mitosis.