Imaging single molecules in live cells in 4+ D (space, time and colors) is crucial for studying various biological processes, especially for observing the behavior of RNA molecules within the nuclear landscape . RNA molecules are known to serve a multitude of tasks such as being templates for protein translation or to act as enzymes for regulating countless reactions in the nucleus . Studying RNA kinetics in living cells can provide new information on RNA function or even human diseases, for instance caused by viruses such as the human immunodeficiency virus (HIV) . A challenge to imaging nuclear RNA function is that the nucleus as a whole undergoes major reformation during the cell cycle  but the time required to step through the sample limits the capability to image large numbers of rapidly moving particles in a 3D space.
- 3D microscopy,
- mRNA trafficking,
- single molecule imaging
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