Time-lapse imaging of nuclear bodies

Fluorescence microscopy is a powerful technique that has become central in the study of the structure and function of biological specimens. This is due in large part to its specifi city and versatility. Although an understanding of structure—typically through high-resolution imaging of fi xed material—has proved animportant tool to understanding function, fl uorescence microscopy also offers a mechanism to interrogate cells in the living state, providing a means to explore dynamic process within the specimen over long time periods at high temporal resolution. The cell nucleus is a highly compartmented environment whose components are often highly motile and in a constant state of fl ux. The ability to monitor the dynamic behavior of nuclear bodies by live-cell imaging provides the researcher with important information regarding underlyingmechanistic processes relating to their formation and maintenance. Two techniques have proved particularly valuable to our study of cellular dynamics and molecular mobility, namely, time-lapse imaging and fl uorescence recovery after photobleaching (FRAP). Time-lapse microscopy allows for qualitative andquantitative analysis of a wide range of events at the cellular and subcellular level. FRAP provides a mechanism to study the mobility of a population of proteins in a range of conditions within discrete areas of the biological specimen. Therefore, fl uorescence microscopy is unique in its ability to provide data at high temporal resolution and in such exquisite detail