Single-image axial localization precision analysis for individual fluorophores.
Single-image axial localization precision analysis for individual fluorophores.
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DOI：
10.1364/oe.20.003057
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发表时间：
2012-01-30
期刊：
影响因子：
3.8
通讯作者：
Wang YM
中科院分区：
文献类型：
--
作者：
DeSantis MC;Zareh SK;Li X;Blankenship RE;Wang YM
生物机制研究需要具有高空间和时间分辨率的单粒子跟踪，这需要具有匹配精度和速度的单荧光团 3D 定位测量。尽管横向定位测量的精度可以通过解析表达式很好地描述，但对于轴向方向，它通常是通过重复位置测量或估计下限来获得的。在这里，我们报告了轴向定位方法的精确表达式，该方法分析单个荧光团强度分布的标准偏差。与横向定位精度一样，该表达式包括可从荧光团的高斯强度分布测量的所有相关实验效应。该表达式完成了对单个荧光团的单图像 3D 定位的精确分析，并将时间分辨率提升至典型的毫秒曝光时间尺度。
Bio-mechanism investigations demand single particle tracking with high spatial and temporal resolutions which require single fluorophore 3D localization measurements with matching precision and speed. Although the precision for lateral-localization measurements is well described by an analytical expression, for the axial direction, it is often obtained by repeating location measurements or by estimating a lower bound. Here, we report a precision expression for an axial-localization method that analyzes the standard deviations of single fluorophores’ intensity profiles. Like the lateral-localization precision, this expression includes all relevant experimental effects measurable from a Gaussian intensity profile of the fluorophore. This expression completes the precision analysis for single-image 3D localization of individual fluorophores and lifts the temporal resolution to the typical exposure timescales of milliseconds.