Colyer Ryan A, Lee Claudia, Gratton Enrico
Department of Biomedical Engineering, University of California, Irvine, USA.
Microsc Res Tech. 2008 Mar;71(3):201-13. doi: 10.1002/jemt.20540.
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.
荧光寿命成像（FLIM）是一种强大的显微镜技术，可通过分子种类或其环境的差异来提供生物及其他系统的对比度。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FLIM（DFD-FLIM）系统的数学模型和物理实现方案，该系统能够提供与时间相关单光子计数方法相当的寿命分辨率。我们的实现方案以相量形式原生提供数据。基于该数学模型，我们进行了误差分析，给出了用于最大化寿命采集质量的精确参数，以及支持这一结论的数据。所提出的DFD-FLIM方法的硬件和软件简化了FLIM的数据采集过程，提供了一种新的数据显示和解释界面，并优化了寿命测定的准确性。
