Pande P, Trivedi C A, Jo J A
Department of Biomedical Engineering, Texas A&M University, 3120 TAMU, College Station, Texas 77843, USA.
Methods Inf Med. 2010;49(5):531-6. doi: 10.3414/ME09-02-0046. Epub 2010 Jul 20.
A novel Fluorescence Lifetime Imaging Microscopy (FLIM) deconvolution method based on the linear expansion of fluorescence decays on a set of orthonormal Laguerre functions was recently proposed. The Laguerre deconvolution method applies linear least-square estimation to estimate the expansion coefficients of all pixel decays simultaneously, performing at least two orders of magnitude faster than the other algorithms. In the original Laguerre FLIM deconvolution implementation, however, the Laguerre parameter α is selected using a heuristic approach, making it unsuitable for online applications.
In this study, we present a fully automated implementation of the Laguerre FLIM deconvolution, whereby the Laguerre parameter α is treated as a free parameter within a nonlinear least-squares optimization scheme.
The performance of this method has been successfully validated on simulated data, and experimental FLIM images of standard fluorescent dyes and endogenous tissue fluorescence.
The main advantage of the proposed method is that it does not require any user intervention for tuning up the deconvolution process. Thus, we believe this method will facilitate the translation of FLIM to online applications, including real-time clinical diagnosis.
最近提出了一种基于荧光衰减在一组正交拉盖尔函数上的线性展开的新型荧光寿命成像显微镜（FLIM）反卷积方法。拉盖尔反卷积方法应用线性最小二乘估计来同时估计所有像素衰减的展开系数，其执行速度比其他算法快至少两个数量级。然而，在原始的拉盖尔FLIM反卷积实现中，拉盖尔参数α是使用启发式方法选择的，这使其不适用于在线应用。
在本研究中，我们提出了拉盖尔FLIM反卷积的全自动实现，其中拉盖尔参数α在非线性最小二乘优化方案中被视为自由参数。
该方法的性能已在模拟数据以及标准荧光染料和内源性组织荧光的实验FLIM图像上成功得到验证。
所提出方法的主要优点是它在反卷积过程的调整中不需要任何用户干预。因此，我们相信该方法将促进FLIM向在线应用的转化，包括实时临床诊断。
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