EXPERIMENTAL CONDITIONS FOR USE OF NITROXIDES IN CELL STUDIES

在细胞研究中使用氮氧化物的实验条件

• 批准号: 6120708
• 负责人: PHILIP D MORSE
• 金额: $ 0.16万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-15 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6120708
• 关键词: animal tissue; biological products; biomedical resource; gas

EPR studies of cellular oxygen and nitroxide metabolism often involve a variety of experimental conditions including different sample preparations, cellular concentrations, and spectrometer settings. Post-processing of the resulting spectra also varies greatly. Here, we show that experimental conditions can affect results often in unexpected ways. We present data on the effects of cell density, type of cell bathing media, and nitroxide concentration in the sample, and power levels and modulation intensity setting of the spectrometer, on oxygen and nitroxide consumption results. We also compare information obtained from simple line height measurements with information obtained from least-squares spectral simulation techniques. Our most consistent results are obtained at low (105 cells/ml or less) cell density, media which alone does not reduce nitroxides, low (0.1mW) power, modulation levels appropriate for good (100:1 or better) signal-to-noise, and lineshape analysis by a fast convolution algorithm combined with Levenberg-Marquard optimization (for determining oxygen and nitroxide concentrations).

细胞氧和氮氧化物代谢的EPR研究 涉及各种实验条件，包括不同的 样品制备、细胞浓度和光谱仪 设置。产生的光谱的后处理也不同 非常好。在这里，我们展示了实验条件可以影响 结果往往是意想不到的。我们提供了有关以下因素影响的数据 细胞密度、细胞洗浴介质类型和氮氧化物浓度 在样本中，以及功率电平和调制强度设置 在光谱仪上，对氧气和氮氧化物的消耗结果。我们 也可以比较从简单的线高测量中获得的信息 利用从最小二乘谱模拟获得的信息 技巧。我们最一致的结果是在LOW(105 细胞/毫升或更少)细胞密度，单独不会降低的培养基数 氮氧化物，低(0.1 mW)功率，调制电平适用于 (100：1或更高)信噪比和线形分析 结合Levenberg-Marquard优化的卷积算法 (用于测定氧气和氮氧化物的浓度)。