Evaluation of global analysis algorithms for single frequency fluorescence lifetime imaging microscopy data
Evaluation of global analysis algorithms for single frequency fluorescence lifetime imaging microscopy data
复制标题
DOI：
10.1046/j.1365-2818.2003.01093.x
复制
发表时间：
2003-01-01
期刊：
影响因子：
--
通讯作者：
Bastiaens, PIH
中科院分区：
文献类型：
--
作者：
Verveer, PJ;Bastiaens, PIH
荧光寿命图像显微镜（FLIM）数据的全局分析可用于获得多指数荧光衰减的精确拟合。特别是，它可以用于拟合单频FLIM数据的双指数衰减，这是传统拟合技术无法实现的。双指数荧光衰减模型可用于定量分析单频FLIM数据的样品，表现出荧光共振能量转移（FRET）。全局分析算法同时拟合在不同实验条件下获得的多个测量值，以获得更高的精度。为了证明双指数模型确实可以拟合到单频数据，我们推导了两个测量的特殊情况的解析解，并使用该解来说明全局分析算法的性质。我们还推导了一种新的全局分析算法，该算法针对单频FLIM数据进行了优化，并证明了它在计算需求方面优于早期的算法。
Global analysis of fluorescence lifetime image microscopy (FLIM) data can be used to obtain an accurate fit of multi-exponential fluorescence decays. In particular, it can be used to fit a bi-exponential decay to single frequency FLIM data, which is not possible with conventional fitting techniques. Bi-exponential fluorescence decay models can be used to analyse quantitatively single frequency FLIM data from samples that exhibit fluorescence resonance energy transfer (FRET). Global analysis algorithms simultaneously fit multiple measurements acquired under different experimental conditions to achieve higher accuracy. To demonstrate that bi-exponential models can indeed be fitted to single frequency data, we derive an analytical solution for the special case of two measurements and use this solution to illustrate the properties of global analysis algorithms. We also derive a novel global analysis algorithm that is optimized for single frequency FLIM data, and demonstrate that it is superior to earlier algorithms in terms of computational requirements.