Abstract
The nucleolus is the hallmark of nuclear compartmentalization and has been shown to exert multiple roles in cellular metabolism besides its main function as the place of ribosomal RNA synthesis and assembly of ribosomes. The nucleolus plays also a major role in nuclear organization as the largest compartment within the nucleus. The prominent structure of the nucleolus can be detected using contrast light microscopy providing an approximate localization of the nucleolus, but this approach does not allow to determine accurately the three-dimensional structure of the nucleolus in cells and tissues. Immunofluorescence staining with antibodies specific to nucleolar proteins albeit very useful is time consuming, normally antibodies recognize their epitopes only within a small range of species and is applicable only in fixed cells. Here, we present a simple method to selectively and accurately label this ubiquitous subnuclear compartment in living cells of a large range of species using a fluorescently labeled cell-penetrating peptide.
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Acknowledgements
We thank all present and past members of the laboratory for their contributions over the years. Robert M. Martin is supported by a fellowship of the Fundação para a Ciência e Tecnologia, Portugal (SFRH/BPD/66611/2009). The laboratory of M. Cristina Cardoso is supported by grants of the German Research Foundation (DFG) and the Federal Ministry of Education and Research (BMBF).
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Martin, R.M., Herce, H.D., Ludwig, A.K., Cardoso, M.C. (2016). Visualization of the Nucleolus in Living Cells with Cell-Penetrating Fluorescent Peptides. In: Németh, A. (eds) The Nucleolus. Methods in Molecular Biology, vol 1455. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3792-9_6
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DOI: https://doi.org/10.1007/978-1-4939-3792-9_6
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