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Suitable for a variety of applications in biology and biomedical research, fluorescence lifetime imaging (FLIM) is a powerful new tool for advanced quantitative analysis. Its high contrast and sensitivity allow for the detection of signals at the single molecule level. After an overview of basic principles of fluorescence, microscopy, and FLIM, this book discusses probes, dynamic and video rate FLIM imaging, laser scanning confocal FLIM, spectral lifetime imaging, nonlinear curve-fitting methods, global FLIM data analysis, and new approaches in data analysis as well as current applications.