Fluorescence microscopy and associated biophysical methods are integral to many areas of biological research including biomedical engineering, cell biology, and molecular biology. This course covers the theory, mechanics, and application of fluorescent microscopy. Students will gain expertise in basic and advanced quantitative fluorescence microscopy in the context of working with living samples. The course topics include sample preparation (immunofluorescence-, dye-, and fluorescent protein-labeling), multidimensional imaging, confocal microscopy, two-photon microscopy, and other advanced imaging techniques. The course will also cover the associated biophysical methods used to probe live cell dynamics such as fluorescence recovery after photobleaching (FRAP), Förster resonance energy transfer (FRET), and fluorescence correlation spectroscopy (FCS). By centering on applications to living samples, students with gain the theoretical background and practical knowledge to design and implement live cell imaging experiments.