Analyzing Membrane Dynamics with Live Cell Fluorescence Microscopy with a Focus on Yeast Mitochondria

With the availability of increasing numbers of fluorescent protein variants and state-of-the-art imaging techniques, live cell microscopy has become a standard procedure in modern cell biology. Fluorescent markers are used to visualize the dynamic processes that take place in living cells, including the behavior of membrane-bound organelles. Here, we provide two examples of how we analyze the membrane dynamics of mitochondria in living yeast cells using wide field and confocal microscopy: (1) Long-term observation of mitochondrial shape changes using mitochondria-targeted fluorescent proteins and (2) monitoring the behavior of individual mitochondria using a mitochondria-targeted version of a photoconvertible fluorescent protein.