Single plane illumination microscopy as a tool for studying nucleome dynamics

Publication date: Available online 23 June 2017 Source:Methods Author(s): Jörg Langowski Single plane illumination microscopy (SPIM) is a new optical method that has become extremely important in recent years. It is based on the formation of a “light slice“ in the specimen in which fluorescently tagged molecules are observed. The spatial resolution is close to that of confocal optics, but without the disadvantages inherent to scanning or high laser irradiation doses. A recent development is light sheet fluctuation microscopy, which exploits the dynamic information contained in the fluorescence intensity fluctuations of each image pixel. Here we review the principles of this method and show some recent appli-cations to the dynamics of transcription factors and chromatin. We show that the dimerization of Fos and Jun proteins is directly linked to their binding to DNA; that nuclear receptor activation changes their intranuclear dynamics; and that the viscoelastic behavior of interphase chromatin strongly depends on the presence of lamin A.
Source: Methods - Category: Molecular Biology Source Type: research