2012
DOI: 10.1111/j.1365-2818.2011.03592.x
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Current SEM techniques for de‐ and re‐construction of centromeres to determine 3D CENH3 distribution in barley mitotic chromosomes

Abstract: SummaryCombined light microscopic (LM) and field emission scanning electron microscopic (FESEM) techniques with FluoroNanogold labelling allowed quantification and high resolution analysis of 3D distribution of the centromere-specific histone H3 variant CENH3 in barley mitotic chromosomes. Chromosomes were investigated with fluorescence LM, conventional FESEM, low-voltage FESEM and combined FIB/FESEM techniques for unprecedented comprehensive analysis to determine chromatin distribution patterns in the centrom… Show more

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Cited by 20 publications
(13 citation statements)
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“…As it is crucial to define the coordinates of a target area for re-localization in SEM we developed a variety of slides with coordinates, successively improved for different demands (Schroeder-Reiter et al 2012). Our aim was to embed cells on slides or cover slips within an ultra-thin resin layer (i) for rapid and precise correlation to LM micrographs and (ii) to allow FIB-milling in any desired direction.…”
Section: Introductionmentioning
confidence: 99%
“…As it is crucial to define the coordinates of a target area for re-localization in SEM we developed a variety of slides with coordinates, successively improved for different demands (Schroeder-Reiter et al 2012). Our aim was to embed cells on slides or cover slips within an ultra-thin resin layer (i) for rapid and precise correlation to LM micrographs and (ii) to allow FIB-milling in any desired direction.…”
Section: Introductionmentioning
confidence: 99%
“…Dual beam focused ion beam/field emission scanning electron microscopy (FIB/FESEM) investigations were performed on a Zeiss Auriga CrossBeam Workstation, a field emission scanning electron microscope equipped with a Gallium ion beam, in-lens, chamber SE and EsB detectors (Carl Zeiss, Germany) as described in Schroeder-Reiter et al [37]. For FIB/FESEM sectioning the specimens were carbon-coated to 10 nm for stability.…”
Section: Methodsmentioning
confidence: 99%
“…However, centromere chromatin does not consist exclusively of CENP-A nucleosomes 2 4 , as canonical histone H3 nucleosomes are also present at the centromere. In all models of centromere chromatin structure in mitotic chromosomes, the CENP-A nucleosomes are found exclusively on the external face of the centromere, whereas the inner centromere contains exclusively H3 nucleosomes 2 , 4 7 . The N-terminal tail of CENP-A differs considerably from that of H3, and between species.…”
Section: Introductionmentioning
confidence: 99%