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The nucleus is the defining feature of eukaryotic cells. It is a highly dynamic, membrane-bound organelle that encloses chromatin and thereby partitions gene transcription from sites of protein translation in the cytoplasm. Major cellular events, including DNA replication, messenger RNA synthesis and processing, and ribosome subunit biogenesis, take place within the nucleus, resulting in a continuous flux of macromolecules into and out of the nucleus through dedicated nuclear pore complexes in the nuclear envelope. Here, we review the impact of new technologies, especially in areas of fluorescence microscopy and proteomics, which are providing major insights into dynamic processes affecting both structure and function within the nucleus.