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Nanoimaging pp 397–416Cite as

Correlative Fluorescence and EFTEM Imaging of the Organized Components of the Mammalian Nucleus

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 950))

Abstract

The cell nucleus contains many distinct subnuclear compartments, domains, and bodies that vary in their composition, structure, and function. While the cellular constituents that occupy the subnuclear regions may be well known, defining the structural details of the molecular assembly of the constituents has been more difficult. A correlative fluorescence and energy-filtering transmission electron microscopy (EFTEM) imaging method has the ability to provide these details. The correlative microscopy method described here allows the tracking of subnuclear structures from specific cells by fluorescence microscopy and then, using electron energy loss imaging in the transmission electron microscope, reveals the ultrastructural features of the nuclear components along with endogenous elemental information that relates directly to the biochemical composition of the structure. The ultrastructural features and composition of well-characterized PML bodies and interchromatin granule clusters are compared to those of ligand-activated glucocorticoid receptor (GR) foci, with GR foci containing fibrogranular nucleic acid-containing features and PML bodies being devoid of nucleic acid.

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Acknowledgments

The EFTEM images were collected with generous support from David Bazett-Jones. Ying Ren helped prepare the samples, and Michael Hendzel and Alioscka Sousa provided helpful critical reading of the manuscript. Maria Aronova and Richard Leapman generously provided advice, and access to multiple EFTEM microscopes. This research was supported in part by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.

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Authors and Affiliations

  1. Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Michael J. Kruhlak

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  1. Michael J. Kruhlak
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Correspondence to Michael J. Kruhlak .

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Editors and Affiliations

  1. Nat. Inst. Biomedical Imaging & Bioengin, National Institutes of Health, South Drive 13, Bethesda, 20892, Maryland, USA

    Alioscka A. Sousa

  2. , National Cancer Institute, National Institutes of Health, Center Drive 10, Bethesda, 20892, USA

    Michael J. Kruhlak

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Kruhlak, M.J. (2013). Correlative Fluorescence and EFTEM Imaging of the Organized Components of the Mammalian Nucleus. In: Sousa, A., Kruhlak, M. (eds) Nanoimaging. Methods in Molecular Biology, vol 950. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-137-0_22

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  • DOI: https://doi.org/10.1007/978-1-62703-137-0_22

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-136-3

  • Online ISBN: 978-1-62703-137-0

  • eBook Packages: Springer Protocols

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