A Phosphorimager for Research in Plant Biology

植物生物系和农业生物技术中心的一组研究人员要求一种多用户磷成像仪。这种仪器通常被称为快速、准确、无胶片的放射自显影系统。它有助于在凝胶或印迹上快速和敏感地定量放射性，而无需闪烁计数或放射自显影。高灵敏度存储荧光粉屏幕捕获实验样品的图像比胶片快十倍。宽线性动态范围的磷光成像仪允许定量检测强弱信号的方式，胶卷根本不能。用微机对数字图像进行编译和分析。我们建议在广泛的应用中使用这种强大的仪器，包括通过凝胶阻滞来定量分析dna -蛋白质相互作用，通过RNA凝胶印迹或RNase保护测定来测量转录物水平，以及酶活性和蛋白质磷酸化的动力学测量。获得磷光成像仪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术是不可能获得的。七名调查人员中有六名目前得到联邦机构竞争性拨款的支持。在所有的情况下，需要磷成像仪分析的拟议实验对于理解植物发病机制、信号转导、膜运输和有丝分裂的分子基础是必不可少的。

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.