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Transmission electron microscopy imaging to analyze chromatin density distribution at the nanoscale level


Fabrice, Tohnyui Ndinyanka; Cherkezyan, Lusik; Ringli, Christoph; Baroux, Célia (2018). Transmission electron microscopy imaging to analyze chromatin density distribution at the nanoscale level. In: Bemer, Marian; Baroux, Célia. Plant Chromatin Dynamics : Methods and Protocols. New York, NY: Springer, 633-651.

Abstract

Transmission electron microscopy (TEM) is used to study the fine ultrastructural organization of cells. Delicate specimen preparation is required for results to reflect the "native" ultrastructural organization of subcellular features such as the nucleus. Despite the advent of high-resolution, fluorescent imaging of chromatin components, TEM still provides a unique and complementary level of resolution capturing chromatin organization at the nanoscale level. Here, we describe the workflow, from tissue preparation, TEM image acquisition and image processing, for obtaining a quantitative description of chromatin density distribution in plant cells, informing on local fluctuations and periodicity. Comparative analyses then allow to elucidate the structural changes induced by developmental or environmental cues, or by mutations affecting specific chromatin modifiers at the nanoscale level. We argue that this approach remains affordable and merits a renewed interest by the plant chromatin community.

Abstract

Transmission electron microscopy (TEM) is used to study the fine ultrastructural organization of cells. Delicate specimen preparation is required for results to reflect the "native" ultrastructural organization of subcellular features such as the nucleus. Despite the advent of high-resolution, fluorescent imaging of chromatin components, TEM still provides a unique and complementary level of resolution capturing chromatin organization at the nanoscale level. Here, we describe the workflow, from tissue preparation, TEM image acquisition and image processing, for obtaining a quantitative description of chromatin density distribution in plant cells, informing on local fluctuations and periodicity. Comparative analyses then allow to elucidate the structural changes induced by developmental or environmental cues, or by mutations affecting specific chromatin modifiers at the nanoscale level. We argue that this approach remains affordable and merits a renewed interest by the plant chromatin community.

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Additional indexing

Item Type:Book Section, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Plant and Microbial Biology
Dewey Decimal Classification:580 Plants (Botany)
Uncontrolled Keywords:Contrast staining; Euchromatin; Heterochromatin; High-pressure freezing; Nanoscale-level chromatin organization; Nucleus; Spatial chromatin density distribution; TEM; Transmission electron microscopy
Language:English
Date:2018
Deposited On:04 Jan 2018 18:23
Last Modified:20 Feb 2018 08:59
Publisher:Springer
Series Name:Methods in Molecular Biology
ISSN:1064-3745
ISBN:978-1-4939-7317-0
OA Status:Closed
Publisher DOI:https://doi.org/10.1007/978-1-4939-7318-7_34
Related URLs:https://doi.org/10.1007/978-1-4939-7318-7 (Publisher)
PubMed ID:29052215

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