Estimation of migration-time and mobility distributions in organelle capillary electrophoresis with statistical-overlap theory.
Estimation of migration-time and mobility distributions in organelle capillary electrophoresis with statistical-overlap theory.
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DOI：
10.1021/ac901982u
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发表时间：
2010-01-01
影响因子：
7.4
通讯作者：
Arriaga, Edgar A.
中科院分区：
文献类型：
--
作者：
Davis, Joe M.;Arriaga, Edgar A.
毛细管电泳（CE）分离细胞器产生大量的窄峰，通常被认为是来自单个颗粒。在本文中，我们表明这并不总是正确的。在这里，我们使用已建立的方法来分区模拟和真实的细胞器CE到恒定的峰密度的区域，然后使用重叠理论来计算每个区域中的峰（单个粒子）的数量。唯一需要的测量值是观察到的峰（最大值）的数量和区域中的峰标准差，以及区域的持续时间。理论发展的估计峰数的精度和阈值饱和度以上的计算是不可取的，由于峰数的波动。理论分析表明，当饱和度在0.2 ~ 1.0之间时，相对精度较好，当饱和度略大于0.5时，相对精度最佳。它还显示阈值饱和度取决于峰值标准差除以区域的持续时间。在细胞器CE的不同区域中估计的峰数的准确性和精确度通过具有恒定和非均匀峰密度的计算机模拟来验证。估计精确到6%。利用不同区域的峰数估计值计算了迁移时间和迁移率分布。这些分布是由峰重叠比计数最大值确定的偏差较小，并提供更正确的措施的细胞器的属性。该程序应用于线粒体CE，其中超过20%的峰被隐藏的峰重叠。
The separation of organelles by capillary electrophoresis (CE) produces large numbers of narrow peaks, which commonly are assumed to originate from single particles. In this paper, we show this is not always true. Here, we use established methods to partition simulated and real organelle CEs into regions of constant peak density and then use statistical-overlap theory to calculate the number of peaks (single particles) in each region. The only required measurements are the number of observed peaks (maxima) and peak standard deviation in the regions, and the durations of the regions. Theory is developed for the precision of the estimated peak number and the threshold saturation above which the calculation is not advisable due to fluctuation of peak numbers. Theory shows that the relative precision is good, when the saturation lies between 0.2 and 1.0, and is optimal when the saturation is slightly greater than 0.5. It also shows the threshold saturation depends on the peak standard deviation, divided by the region’s duration. The accuracy and precision of peak numbers estimated in different regions of organelle CEs are verified by computer simulations having both constant and non-uniform peak densities. The estimates are accurate to 6%. The estimated peak numbers in different regions are used to calculate migration-time and electrophoretic-mobility distributions. These distributions are less biased by peak overlap than ones determined by counting maxima and provide more correct measures of the organelle properties. The procedure is applied to a mitochondrial CE, in which over 20% of peaks are hidden by peak overlap.
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影响因子：
1.7
作者：
Dondi, F;Pietrogrande, MC;Felinger, A
通讯作者：
Felinger, A
影响因子：
4.3
作者：
Johnson, Ryan D.;Navratil, Marian;Arriaga, Edgar A.
通讯作者：
Arriaga, Edgar A.
影响因子：
7.4
作者：
CHENG, YF;WU, SL;DOVICHI, NJ
通讯作者：
DOVICHI, NJ
影响因子：
4.1
作者：
Davis, Joe M.;Arriaga, Edgar A.
通讯作者：
Arriaga, Edgar A.
影响因子：
7.4
作者：
Felinger, A
通讯作者：
Felinger, A