Stopped flow spectrofluorometer

在由该提案的9名共同研究者进行的10个NIH支持的核心项目中，荧光光谱法为蛋白质构象转变的动态方面提供了一个独特的窗口，伴随着肽-膜相互作用和插入，核酸-蛋白质相互作用，以及与其蛋白质或核酸靶点结合的药物分子的活性构象的研究。我们寻求获得最先进的荧光光谱仪，以支持和加强这些研究计划。这种仪器将取代老化的荧光光谱仪，现在很容易崩溃。此外，该系统由一台过时的、不符合Y2K标准的计算机控制，而且无法获得更换零件。现有的仪器已经作为共享设施的一部分运行了大约6年;在这段时间里，它已经为来自该大学8个部门和4个部门的20多名研究人员提供了服务，并随着时间的推移，为20个或更多NIH支持的项目提供了支持。将购买的仪器将配备荧光各向异性测量，并将有一个停流附件，以支持那些需要快速动力学测量的项目。二极管阵列检测器和双单色器将能够在短时间内快速采集整个光谱。吸光度测量将是可能的，使监测自组装反应或聚集。该工具将支持由7名NIH支持的研究者和另外两名核心研究者组成的主要小组。其中既有广泛的，可证明的需求，以及荧光和停流技术的经验。过渡到现代化的、可靠的、计算机控制的数据采集仪器，不仅可以使现有仪器不可行的实验方法成为可能，而且还可以通过减少修理的停机时间和用于为过时的计算机系统寻找功能部件的相当持续的时间消耗来提高生产率。总的来说，广泛的计算机控制的数据采集，快速数据采集和停流荧光代表的能力，将大大提高现有的NIH 0支持的项目。

In ten NIH-supported core projects conducted by the 9 co-investigators of this proposal, fluorescence spectroscopy provides a unique window into dynamic aspects of protein conformational transitions accompanying peptide-membrane interaction and insertion, nucleic acid- protein interaction, and investigations of the active conformation of drug molecules bound to their protein or nucleic acid targets. We seek to acquire a state-of-the-art fluoresc4ence spectrometer to support and enhance these research programs. This instrument would replace an aging fluorescence spectrometer that is now prone to breakdown. Also, it is controlled by an obsolete, non-Y2K-compliant computer for which replacement parts are unavailable. The existing instrument has been operated as part of a shared facility for approximately 6 years; in that time it has served more than 20 investigators from in 8 departments and 4 divisions of this University, and over time has provided support to 20 or more NIH-supported projects. The instrument to be purchased will be equipped to enable fluorescence anisotropy measurements, and will have a stopped-flow accessory to support those projects requiring rapid kinetics measurements. Diode array detectors and dual monochromators will enable rapid acquisition of entire spectra at short time scales. Absorbance measurements will be possible, enabling the monitoring of self-assembly reactions or aggregation. This instrument will support a principal group of 7 NIH-supported investigators and two additional core investigators and two additional core investigators. Among them there is both have extensive, demonstrable need, as well as experience with fluorescence and stopped-flow techniques. The transition to modern, reliable instrumentation with computer-controlled data acquisition will not only make possible experimental approaches that are not feasible with the existing instrument, but also enhance productivity by reducing down time for repairs and the fairly constant drain of time devoted to scavenging functional parts for the obsolete computer system. Over all, extensive computer-controlled data acquisition, rapid data acquisition, and stopped-flow fluorescence represent capabilities that will enhance greatly the existing NIH0-supported projects.