Imaging green fluorescent protein fusions in living fission yeast cells |
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Authors: | Tran P T Paoletti Anne Chang Fred |
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Affiliation: | University of Pennsylvania, Department of Cell and Developmental Biology, 421 Curie Blvd.-BRB2/3 Room 1009, Philadelphia, PA 19104, USA. |
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Abstract: | The use of green fluorescent protein (GFP) fusions as biosensors for examining protein localization and dynamics has revolutionized cell biology. Here, we describe the methods developed for imaging of GFP-fusions in the fission yeast Schizosaccharomyces pombe using fluorescence microscopy, with a focus on the use of time-lapse imaging to analyze the dynamics of microtubules. We discuss the considerations in fluorescence microscopy, cell preparation, data acquisition, and image analysis appropriate for analysis of living cells. |
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