ArticlePloS one2010
Condensed mitotic chromosome structure at nanometer resolution using PALM and EGFP- histones.
Article in PloS one, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed, 87 citations in OpenAlex.
- Spectroscopic Super-resolution Imaging of DNA Molecules using Intrinsic Contrast.Journal of visualized experiments : JoVE · 2026Article
- ChromEMT: visualizing and reconstructing chromatin ultrastructure and 3D organization in situ.Nature protocols · 2025Review
- Enabling real-time reconstruction for large field-of-view single-molecule localization microscopy using discrete field-dependent point-spread function.Biomedical optics express · 2025Article
- Insight into chromatin compaction and spatial organization in rice interphase nuclei.Frontiers in plant science · 2024Article
- Review
- Advanced imaging techniques for tracking drug dynamics at the subcellular level.Advanced drug delivery reviews · 2023Review
- Mitotic chromosomes.Seminars in cell & developmental biology · 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
- Seeing chromosome structure reveals its function.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2021Article
- Cryo-nanoscale chromosome imaging-future prospects.Biophysical reviews · 2020Review
- A General Mechanism of Green-to-Red Photoconversions of GFP.Frontiers in molecular biosciences · 2020Article
- Asymmetrical localization of Nup107-160 subcomplex components within the nuclear pore complex in fission yeast.PLoS genetics · 2019Article
- Use of 3D imaging for providing insights into high-order structure of mitotic chromosomes.Chromosoma · 2019Review
- Challenges and guidelines toward 4D nucleome data and model standards.Nature genetics · 2018Review
- Switchable Fluorophores for Single-Molecule Localization Microscopy.Chemical reviews · 2018Review
- Visualization of chromosome condensation in plants with large chromosomes.BMC plant biology · 2017Article
- ChromEMT: Visualizing 3D chromatin structure and compaction in interphase and mitotic cells.Science (New York, N.Y.) · 2017Article
- Stochastic fluorescence switching of nucleic acids under visible light illumination.Optics express · 2017Article
- Superresolution imaging of nanoscale chromosome contacts.Scientific reports · 2017Article
- Borna Disease Virus Assembles Porous Cage-like Viral Factories in the Nucleus.The Journal of biological chemistry · 2016Article
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
Abstract
Photoactivated localization microscopy (PALM) and related fluorescent biological imaging methods are capable of providing very high spatial resolutions (up to 20 nm). Two major demands limit its widespread use on biological samples: requirements for photoactivatable/photoconvertible fluorescent molecules, which are sometimes difficult to incorporate, and high background signals from autofluorescence or fluorophores in adjacent focal planes in three-dimensional imaging which reduces PALM resolution significantly. We present here a high-resolution PALM method utilizing conventional EGFP as the photoconvertible fluorophore, improved algorithms to deal with high levels of biological background noise, and apply this to imaging higher order chromatin structure. We found that the emission wavelength of EGFP is efficiently converted from green to red when exposed to blue light in the presence of reduced riboflavin. The photon yield of red-converted EGFP using riboflavin is comparable to other bright photoconvertible fluorescent proteins that allow <20 nm resolution. We further found that image pre-processing using a combination of denoising and deconvolution of the raw PALM images substantially improved the spatial resolution of the reconstruction from noisy images. Performing PALM on Drosophila mitotic chromosomes labeled with H2AvD-EGFP, a histone H2A variant, revealed filamentous components of ∼70 nm. This is the first observation of fine chromatin filaments specific for one histone variant at a resolution approximating that of conventional electron microscope images (10-30 nm). As demonstrated by modeling and experiments on a challenging specimen, the techniques described here facilitate super-resolution fluorescent imaging with common biological samples.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.