Acquisition of a Spectrofluorometer

技术的重大进步使得监测活细胞内离子活动的快速变化成为可能。该技术首先将不同离子（钙、氢、氯）的荧光染料加载到细胞中。离子浓度的变化是通过监测保存在荧光分光光度计内的细胞的荧光来测量的。这种涉及细胞群的基本技术可以扩展到使用适当的硬件和显微镜分析单细胞。通过微型计算机重建数字化图像，可以对离子浓度变化的细胞进行二维和三维分析。目前，在赖特州立大学，有多达8名独立的研究人员使用相关的荧光技术，并在同一台仪器上共享时间。现有的仪器除了被大量使用外，还有某些缺点。然而，已经获得了足够重要的初步数据，促使五名研究人员根据仪器仪表计划申请资金，购买另一台具有双光束激发、光电二极管阵列检测、单细胞（显微镜）和成像能力的荧光光谱仪。这五名研究人员的研究计划涉及测量不同细胞类型中的钙、pH和氯化物，包括星形胶质细胞、血管平滑肌和人类二倍体成纤维细胞。此外，所要求的荧光光谱仪将用于涉及膜融合的研究，例如在胞吐或病毒入侵期间发生的膜融合。

Major advances in technology have made it possible to monitor rapid changes in intracellular ion activities in living cells. the technique involves first loading cells with fluorescent dyes specific for different ions (Ca , H , Cl ). Changes in ion concentration are then measured by monitoring fluorescence from the cells maintained inside the cuvette of a spectrofluorometer. this basic technique involving a population of cells can be extended to analysis of single cells with the appropriate hardware and microscope. Reconstruction of a digitized image by a mini-computer allows 2 and 3-D analysis of cells undergoing changes in ion concentrations. Currently, at Wright State University, there are as many as eight independent investigators using related fluorescent techniques and sharing time on a single instrument. The available instrument has certain drawbacks as well as being heavily used. However, enough significant preliminary data has been obtained to prompt five researchers to request funds under the Instrumentation Program to purchase another spectrofluorometer with dual beam excitation, photodiode array detection, single cell (microscopic) and imaging capabilities. The research proposal of these five investigators involve the measurement of calcium, pH, and chloride in diverse cell types, including astrocytes, vascular smooth muscle, and human diploid fibroblasts. In addition, the requested spectrofluorometer will be used for studies involving membrane fusion such as that which occurs during exocytosis or viral invasion.