FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye.
FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye.
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DOI：
10.1371/journal.pone.0131640
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Haueisen J
中科院分区：
文献类型：
--
作者：
Klemm M;Schweitzer D;Peters S;Sauer L;Hammer M;Haueisen J
荧光寿命成像检眼镜（FLIO）是一种测量眼底内源性荧光团在体自发荧光强度衰减的新技术。在这里，我们提出了一个软件包，称为FLIM eXplorer（FLIMAX）分析FLIO数据。具体来说，我们引入了一种新的自适应合并方法作为空间分辨率和每个像素所需光子数之间的最佳权衡。我们还扩展了现有的衰减模型（多指数，拉伸指数，光谱全局分析，不完全衰减），以考虑眼睛的分层结构，并提出了一种方法来校正晶体透镜荧光对视网膜荧光的影响。随后，将Holm-Bonferroni方法应用于FLIO测量，以允许基于荧光寿命参数对患者和对照组进行组比较。新方法的性能进行了评估，在五个实验。具体而言，我们评估了糖尿病患者的静态和自适应分箱，我们比较了健康志愿者的不同衰减模型，并在糖尿病患者和对照组之间进行了组比较。一个概述的可视化能力和静态和自适应分箱的比较显示了黄斑裂孔患者。FLIMX的荧光寿命成像显微镜的适用性显示在神经节细胞层的猪视网膜样品，通过激光扫描显微镜使用双光子激发。
Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique for measuring the in vivo autofluorescence intensity decays generated by endogenous fluorophores in the ocular fundus. Here, we present a software package called FLIM eXplorer (FLIMX) for analyzing FLIO data. Specifically, we introduce a new adaptive binning approach as an optimal tradeoff between the spatial resolution and the number of photons required per pixel. We also expand existing decay models (multi-exponential, stretched exponential, spectral global analysis, incomplete decay) to account for the layered structure of the eye and present a method to correct for the influence of the crystalline lens fluorescence on the retina fluorescence. Subsequently, the Holm-Bonferroni method is applied to FLIO measurements to allow for group comparisons between patients and controls on the basis of fluorescence lifetime parameters. The performance of the new approaches was evaluated in five experiments. Specifically, we evaluated static and adaptive binning in a diabetes mellitus patient, we compared the different decay models in a healthy volunteer and performed a group comparison between diabetes patients and controls. An overview of the visualization capabilities and a comparison of static and adaptive binning is shown for a patient with macular hole. FLIMX’s applicability to fluorescence lifetime imaging microscopy is shown in the ganglion cell layer of a porcine retina sample, obtained by a laser scanning microscope using two-photon excitation.