Stopped flow spectrofluorometer

在NIH支持的10个核心项目中，荧光光谱学提供了一个独特的窗口，可以了解伴随着多肽-膜相互作用和插入、核酸-蛋白质相互作用的蛋白质构象转变的动态方面，以及研究结合到蛋白质或核酸靶标上的药物分子的活性构象。我们寻求获得一台最先进的荧光光谱仪来支持和加强这些研究计划。这台仪器将取代现在容易出现故障的老化的荧光光谱仪。此外，它是由一台过时的、不符合Y2K标准的计算机控制的，没有替换部件可用。现有的仪器作为一个共享设施的一部分已经运行了大约6年；在此期间，它为来自该大学8个系和4个部门的20多名调查人员提供了服务，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成能够进行荧光各向异性测量，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。二极管阵列探测器和双单色仪将能够在短时间尺度上快速获取整个光谱。吸光度测量将成为可能，从而能够监测自组装反应或聚集。该工具将支持由7名国家卫生研究院支持的调查人员组成的主要小组，以及另外两名核心调查人员和两名额外的核心调查人员。其中既有广泛的、明显的需求，也有荧光和停流技术的经验。向具有计算机控制的数据采集的现代、可靠仪器的过渡不仅将使现有仪器不可行的实验方法成为可能，而且还将通过减少维修的停机时间和用于清理过时计算机系统的功能部件的相当持续的时间来提高生产率。总体而言，广泛的计算机控制数据采集、快速数据采集和停流荧光表示的能力将极大地增强现有的NIH0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.