CE RAMAN SPECTROSCOPY OF ATP AND OTHER NEUROTRANSMITTERS

描述：普通拉曼光谱和表面增强拉曼光谱 光谱(SERS)将被用作在线检测技术 用毛细管预浓缩的核糖核苷酸和其他神经化学物质 电泳法。拉曼光谱将允许识别和 细胞培养和萃取物中化合物的定量。 等速电泳法，如果需要的话，随后的电泳法将用于 10~(-6)~10~(-4)M的预浓缩化合物用于SERS和10~(-2)M的分析 10-1M浓度，用于正常拉曼光谱分析。 将采用常规的毛细管和微细加工的毛细管。 表面增强拉曼散射系统将采用光刻制造的银 微电极，它可以放置在毛细管的出口处 无场区域。在微细加工的毛细管中，电极系统 将被整合到分离通道中。 当地设计的共焦表观照明微探头光学系统将 用于拉曼光谱。一台高通量光谱仪， 电荷耦合探测器和2瓦532 nm固体激光器将完成 拉曼光谱系统。Gran-Schmidt正交化过程 并将使用其他统计测试来确定和量化目标 初始浓度低至10-9M的化合物合成试验 混合的核糖核苷酸将用于开发和测试 普通拉曼光谱系统。当系统的初始开发是 完成后，将演示在复杂生物基质中的性能 通过测量池塘蜗牛巨大的多巴胺神经元中的谷氨酸。 该系统将被用来研究三磷酸腺苷的水解动力学。 在培养的嗜铬细胞和鸡脑胚胎提取液中。目前， 只有快速降解的三磷酸腺苷才能方便地进行测量和工程 预计会产生比现在更详细和更准确的放映率 可用。此外，三磷酸腺苷/腺苷比率将在相同的 矩阵。最后，该系统将被用于测量c-AMP在PC12中的释放 细胞。

DESCRIPTION: Normal Raman spectroscopy and surface-enhanced Raman spectroscopy (SERS) will be used as on-line detection techniques for ribonucleotides and other neurochemicals preconcentrated by capillary electrophoresis. Raman spectroscopy will allow for identification and quantification of compounds from cell cultures and extracts. Isotachophoresis, with subsequent electrophoresis if needed will be used to preconcentrate compounds to 10-6 - 10-4 M for analysis by SERS and 10-2 - 10-1 M concentration for analysis by normal Raman spectroscopy. Conventional capillaries and microfabricated capillaries will be employed. The SERS system will employ lithographically fabricated silver micro-electrodes, which can be placed at the exit of a capillary in a field-free region. In microfabricated capillaries, the electrode system will be integrated into the separation channel. A locally designed confocal epi-illumination microprobe optical system will be used for Raman spectroscopy. A high throughput spectrograph, charge-coupled detector and a 2 Watt 532 nm solid-state laser will complete the Raman spectroscopy system. The Gran-Schmidt orthogonalization procedure and other statistical tests will be used to identify and quantify target compounds at initial concentrations as low as 10-9 M. Synthetic test mixtures of ribonucleotides will be used for development and testing of the normal Raman spectroscopy system. When initial development of the system is completed, performance in a complex biological matrix will be demonstrated by glutamate measurements in the giant dopamine neuron of the pond snail. The system will be used to study hydrolysis kinetics of ATP to ADP and AMP in cultured chromaffin cells and in chick brain embryo extracts. Presently, only the rapidly hydrolyzed ATP can be conveniently measured and the project is expected to yield more detailed and accurate release rates than are now available. Additionally, ATP/adenosine ratios will be measured in the same matrices. Finally, the system will be used to measure c-AMP release in PC12 cells.