Electron paramagnetic resonance oximetry as a quantitative method to measure cellular respiration: A consideration of oxygen diffusion interference
Electron paramagnetic resonance oximetry as a quantitative method to measure cellular respiration: A consideration of oxygen diffusion interference
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DOI：
10.1529/biophysj.106.090175
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发表时间：
2006-12-01
影响因子：
3.4
通讯作者：
Ilangovan, Govindasamy
中科院分区：
文献类型：
--
作者：
Presley, Tennille;Kuppusamy, Periannan;Ilangovan, Govindasamy
电子顺磁共振（EPR）血氧测定法正被广泛用于测量细胞、线粒体和亚线粒体颗粒的耗氧量。然而，需要在数据分析方面进一步改进这项技术，以将其用作定量工具。在这里，我们提出了一种新的方法，用于定量分析细胞呼吸EPR血氧饱和度。悬浮液中细胞的耗氧过程被观察到有三个不同的区域：在较高pO（2）范围内的pO（2）非依赖性呼吸，在低pO（2）范围内的pO（2）依赖性呼吸，以及在非常低的pO（2）值下pO（2）没有变化的静态平衡。这里的方法使人们能够综合分析所有三个区域-其中考虑每个细胞周围的O-2扩散区域的进展，它们在时间内的重叠，以及它们对测量的pO（2）数据的潜在影响。所获得的结果同意与以前建立的方法，如高分辨率呼吸测量。此外，它也证明了如何扩散限制可以取决于细胞密度和消耗率。总之，新的方法建立了一个更准确和有意义的模型来评估EPR血氧饱和度数据的细胞呼吸，量化相关参数使用EPR血氧饱和度。
Electron paramagnetic resonance (EPR) oximetry is being widely used to measure the oxygen consumption of cells, mitochondria, and submitochondrial particles. However, further improvement of this technique, in terms of data analysis, is required to use it as a quantitative tool. Here, we present a new approach for quantitative analysis of cellular respiration using EPR oximetry. The course of oxygen consumption by cells in suspension has been observed to have three distinct zones: pO(2)-independent respiration at higher pO(2) ranges, pO(2)-dependent respiration at low pO(2) ranges, and a static equilibrium with no change in pO2 at very low pO2 values. The approach here enables one to comprehensively analyze all of the three zones together - where the progression of O-2 diffusion zones around each cell, their overlap within time, and their potential impact on the measured pO(2) data are considered. The obtained results agree with previously established methods such as high-resolution respirometry measurements. Additionally, it is also demonstrated how the diffusion limitations can depend on cell density and consumption rate. In conclusion, the new approach establishes a more accurate and meaningful model to evaluate the EPR oximetry data on cellular respiration to quantify related parameters using EPR oximetry.