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Provides a fundamental understanding of the underlying principles and cell biological applications of fluorescence microscopy, using lectures and in-class activities. Emphasizes high-resolution widefield and confocal fluorescence imaging (fixed and live cells), with topics ranging from immunofluorescence, fluorescent probes, 3D and time-lapse imaging to advanced techniques for measuring dynamics (e.g., FRAP, FLIP) and molecular interactions (e.g., FRET, BiFC), and fluorescence lifetime imaging microscopy (FLIM). Introduces image processing and single-cell resolved quantitative image analysis using open-source software. Presents super-resolution imaging techniques and emerging technologies. Offers practical examples with a focus on organelle biology, cell signaling, and infectious disease.