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June 4, 2018: Hilmar Strickfaden, University of Alberta, Edmonton

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Talk in the biologic colloquium: "Applying Electron Spectroscopic Imaging to answer questions in Nuclear Architecture and Epigenetics"

What
  • Biologisches Kolloquium
When Jun 06, 2018
from 04:15 PM to 05:30 PM
Where Lecture Hall, Zoology, Hauptstr. 1
Contact Name
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Abstract:

Transmission electron microscopy (TEM) has contributed profoundly to the understanding of the structural organization of cells at the nanometer scale. Lack of colors and specific staining methods, however, limits the capacity of TEM for structural discrimination to structures with differences in contrast. This limitation makes it difficult to map the distribution of different biomolecules with similar contrast, such as proteins, RNA and DNA, simultaneously within the same ultrathin section. In order to overcome these problems, we use a method, called Electron Spectroscopic Imaging (ESI) that allows us to map the distribution of elements, such as phosphorus and nitrogen. Although both proteins and nucleic acids contain these elements, they differ profoundly with respect to their ratios to allow their specific discrimination. In addition to elements, which occur naturally in cells, it has become possible to augment ESI by introducing other elements as elemental tags, which allow the discrimination of specific biomolecules, similar to the targeting of structures with fluorescent dyes in fluorescence microscopy.1 The resulting distribution maps yield ESI-images that can be false colored and superimposed on each other to study the interplay between chromatin, the interchromatin space and nuclear bodies. Correlative microscopy allows the sequential studies of the same cell with super-resolved fluorescence microscopy and ESI. Based on this evidence we demonstrate that the functional organization of the mammalian cell nucleus is composed of co-aligned, functionally integrated active and inactive nuclear compartments.2,3

  1. Strickfaden H, Xu ZZ, Hendzel MJ Visualization of miniSOG Tagged DNA Repair Proteins in Combination with Electron Spectroscopic Imaging (ESI). J Vis Exp 2015 Sep 24;(103).
  2. Cremer T, Cremer M, Hübner B, Strickfaden H, Smeets D, Popken J, Sterr M, Markaki Y, Rippe K, Cremer C (2015) The 4D nucleome: Evidence for a dynamic nuclear landscape based on co-aligned active and inactive nuclear compartments. FEBS Lett 589(20 Pt A):2931-2943.
  3. Cremer C, Szczurek A, Strickfaden H, Birk U, Cremer M, Cremer T (2018) Der Zellkern – eine Stadt in der Zelle, Teil 2. Biologie in unserer Zeit 48 (1) 45-53.

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