Acquisition of a Spectrofluorometer

技术的重大进步使监测活细胞内离子活动的快速变化成为可能。这项技术首先在细胞中加入针对不同离子(钙、氢、氯)的荧光染料。然后，通过监测荧光分光光度计试管内细胞的荧光来测量离子浓度的变化。通过适当的硬件和显微镜，这种涉及细胞群体的基本技术可以扩展到对单个细胞的分析。用微型计算机重建数字化图像可以对经历离子浓度变化的细胞进行2-D和3-D分析。目前，在莱特州立大学，多达8名独立调查人员使用相关的荧光技术，在一台仪器上共享时间。现有的仪器有一定的缺点，而且被大量使用。然而，已经获得了足够多的重要初步数据，促使五名研究人员根据仪器方案申请资金，以购买另一台具有双光束激发、光电二极管阵列检测、单电池(显微镜)和成像能力的光谱荧光仪。这五名研究人员的研究建议涉及测量不同类型细胞中的钙、pH和氯，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，所要求的荧光分光光度计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.