A novel fluorescence lifetime imaging system that optimizes photon efficiency
A novel fluorescence lifetime imaging system that optimizes photon efficiency
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DOI：
10.1002/jemt.20540
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发表时间：
2008-03-01
影响因子：
2.5
通讯作者：
Gratton, Enrico
中科院分区：
文献类型：
--
作者：
Colyer, Ryan A.;Lee, Claudia;Gratton, Enrico
荧光寿命成像是一种强大的显微技术，用于通过分子物种或其环境的差异来提供生物和其他系统的对比。然而，设备成本和数据分析的复杂性限制了FLIM的应用。我们提出了一种低成本数字频域薄膜(DFD-FLIM)系统的数学模型和物理实现，该系统可以提供与时间相关单光子计数方法相当的寿命分辨率。我们的实现以相量的形式在本地提供数据。在数学模型的基础上，我们给出了误差分析，给出了最大化寿命获取质量的精确参数，以及支持这一结论的数据。该方法的硬件和软件简化了薄膜的数据采集过程，为数据的显示和解释提供了新的界面，并优化了寿命测定的准确性。
Fluorescence lifetime imaging (FLIM) is a powerful microscopy technique for providing contrast of biological and other systems by differences in molecular species or their environments. However, the cost of equipment and the complexity of data analysis have limited the application of FLIM. We present a mathematical model and physical implementation for a low cost digital frequency domain FLIM (DFD-FLIM) system, which can provide lifetime resolution with quality comparable to time-correlated single photon counting methods. Our implementation provides data natively in the form of phasors. On the basis of the mathematical model, we present an error analysis that shows the precise parameters for maximizing the quality of lifetime acquisition, as well as data to support this conclusion. The hardware and software of the proposed DFD-FLIM method simplifies the process of data acquisition for FLIM, presents a new interface for data display and interpretation, and optimizes the accuracy of lifetime determination.