CE RAMAN SPECTROSCOPY OF ATP AND OTHER NEUROTRANSMITTERS

ATP 和其他神经递质的 CE 拉曼光谱

• 批准号: 2519057
• 负责人: MICHAEL DAVID MORRIS
• 金额: $ 15.71万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-27 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2519057
• 关键词: PC12 cells; Raman spectrometry; adenosine triphosphate; chemical kinetics; chick embryo; chromaffin cells; cyclic AMP; hydrolysis; nucleotide metabolism

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.

描述：正常拉曼光谱和表面增强拉曼光谱