ArticleChromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology2020
Prospects and limitations of expansion microscopy in chromatin ultrastructure determination.
Article in Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed.
- Plants in the expansion era: recent advances in expansion microscopy in plant systems.Journal of experimental botany · 2025Review
- Analytic Fourier ptychotomography for aberration-free and high-resolution volumetric refractive index imaging.Nature communications · 2025Article
- Super-resolution expansion microscopy in plant roots.The Plant cell · 2025Article
- ExPOSE: a comprehensive toolkit to perform expansion microscopy in plant protoplast systems.The Plant journal : for cell and molecular biology · 2025Article
- Proximity labeling expansion microscopy (PL-ExM) evaluates interactome labeling techniques.Journal of materials chemistry. B · 2024Article
- Expanding boundaries - a cell biologist's guide to expansion microscopy.Journal of cell science · 2024Review
- Combining single-molecule and expansion microscopy in fission yeast to visualize protein structures at the nanostructural level.Open biology · 2024Article
- Protein and lipid expansion microscopy with trypsin and tyramide signal amplification for 3D imaging.Scientific reports · 2023Article
- Proximity Labeling Expansion Microscopy (PL-ExM) resolves structure of the interactome.bioRxiv : the preprint server for biology · 2023Article
- Expansion microscopy: A chemical approach for super-resolution microscopy.Current opinion in structural biology · 2023Review
- Light-sheets and smart microscopy, an exciting future is dawning.Communications biology · 2023Review
- Super-resolution microscopy reveals the number and distribution of topoisomerase IIα and CENH3 molecules within barley metaphase chromosomes.Chromosoma · 2023Article
- Nanoscale fluorescence imaging of biological ultrastructure via molecular anchoring and physical expansion.Nano convergence · 2022Review
- Imaging plant cells and organs with light-sheet and super-resolution microscopy.Plant physiology · 2022Article
- Expansion microscopy facilitates quantitative super-resolution studies of cytoskeletal structures in kinetoplastid parasites.Open biology · 2021Article
- Understanding 3D genome organization by multidisciplinary methods.Nature reviews. Molecular cell biology · 2021Review
- Current and future advances in fluorescence-based visualization of plant cell wall components and cell wall biosynthetic machineries.Biotechnology for biofuels · 2021Review
- Contribution of advanced fluorescence nano microscopy towards revealing mitotic chromosome structure.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2021Review
- Challenges of Using Expansion Microscopy for Super-resolved Imaging of Cellular Organelles.Chembiochem : a European journal of chemical biology · 2021Article
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6 authors.
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Abstract
Expansion microscopy (ExM) is a method to magnify physically a specimen with preserved ultrastructure. It has the potential to explore structural features beyond the diffraction limit of light. The procedure has been successfully used for different animal species, from isolated macromolecular complexes through cells to tissue slices. Expansion of plant-derived samples is still at the beginning, and little is known, whether the chromatin ultrastructure becomes altered by physical expansion. In this study, we expanded isolated barley nuclei and compared whether ExM can provide a structural view of chromatin comparable with super-resolution microscopy. Different fixation and denaturation/digestion conditions were tested to maintain the chromatin ultrastructure. We achieved up to ~4.2-times physically expanded nuclei corresponding to a maximal resolution of ~50-60 nm when imaged by wild-field (WF) microscopy. By applying structured illumination microscopy (SIM, super-resolution) doubling the WF resolution, the chromatin structures were observed at a resolution of ~25-35 nm. WF microscopy showed a preserved nucleus shape and nucleoli. Moreover, we were able to detect chromatin domains, invisible in unexpanded nuclei. However, by applying SIM, we observed that the preservation of the chromatin ultrastructure after the expansion was not complete and that the majority of the tested conditions failed to keep the ultrastructure. Nevertheless, using expanded nuclei, we localized successfully centromere repeats by fluorescence in situ hybridization (FISH) and the centromere-specific histone H3 variant CENH3 by indirect immunolabelling. However, although these repeats and proteins were localized at the correct position within the nuclei (indicating a Rabl orientation), their ultrastructural arrangement was impaired.
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