Evidence map›Paper›PMID 39613943›Full record

ReviewNature protocols2025

ChromEMT: visualizing and reconstructing chromatin ultrastructure and 3D organization in situ.

Horng D Ou, Sebastien Phan, Thomas J Deerinck, Akiko Inagaki, Mark H Ellisman, Clodagh C O'Shea

Abstract readReview
In one paragraph

Review in Nature protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Article
  2. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Horng D OuMolecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA.ORCID 0000-0002-1769-9730
Sebastien PhanNational Center for Microscopy and Imaging Research, University of California San Diego, La Jolla, CA, USA.
Thomas J DeerinckNational Center for Microscopy and Imaging Research, University of California San Diego, La Jolla, CA, USA.
Akiko InagakiMolecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA.ORCID 0009-0001-2208-8512
Mark H EllismanNational Center for Microscopy and Imaging Research, University of California San Diego, La Jolla, CA, USA. mellisman@ucsd.edu.
Clodagh C O'SheaMolecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA. oshea@salk.edu.ORCID 0000-0002-4872-9066

Funding

Viral Vector Core (VVC)P30CA014195 · NCI · SALK INSTITUTE FOR BIOLOGICAL STUDIES · PI Ye Zheng · 1985 to 2026
$82.8M
IVEM and Image Analysis ResourceP41GM103412 · NIGMS · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI ELLISMAN, MARK H · 2012 to 2019
$17.9M
Imaging local and global chromatin structure as a 3D continuum within the nucleusU01EB021247 · NIBIB · SALK INSTITUTE FOR BIOLOGICAL STUDIES · PI ELLISMAN, MARK H, O'SHEA, CLODAGH · 2015 to 2017
$1.8M
Imaging local and global chromatin structure as a 3D continuum within the nucleusU01DA047731 · NIDA · SALK INSTITUTE FOR BIOLOGICAL STUDIES · PI ELLISMAN, MARK H, O'SHEA, CLODAGH · 2018 to 2019
$1.5M
NCI NIH HHS P30 CA014195NIBIB NIH HHS U01 EB021247NIDA NIH HHS U01 DA047731NIGMS NIH HHS P41 GM103412U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS) GM103412U.S. Department of Health & Human Services | NIH | National Institute on Drug Abuse (NIDA) U01DA047731
6 · The paper itself

Abstract

Structure determines function. The discovery of the DNA double-helix structure revealed how genetic information is stored and copied. In the mammalian cell nucleus, up to two meters of DNA is compacted by histones to form nucleosome/DNA particle chains that form euchromatin and heterochromatin domains, chromosome territories and mitotic chromosomes upon cell division. A critical question is what are the structures, interactions and 3D organization of DNA as chromatin in the nucleus and how do they determine DNA replication timing, gene expression and ultimately cell fate. To visualize genomic DNA across these different length scales in the nucleus, we developed ChromEMT, a method that selectively enhances the electron density and contrast of DNA and interacting nucleosome particles, which enables nucleosome chains, chromatin domains, chromatin ultrastructure and 3D organization to be imaged and reconstructed by using multi-tilt electron microscopy tomography (EMT). ChromEMT exploits a membrane-permeable, fluorescent DNA-binding dye, DRAQ5, which upon excitation drives the photo-oxidation and precipitation of diaminobenzidine polymers on the surface of DNA/nucleosome particles that are visible in the electron microscope when stained with osmium. Here, we describe a detailed protocol for ChromEMT, including DRAQ5 staining, photo-oxidation, sample preparation and multi-tilt EMT that can be applied broadly to reconstruct genomic DNA structure and 3D interactions in cells and tissues and different kingdoms of life. The entire procedure takes ~9 days and requires expertise in electron microscopy sample sectioning and acquisition of multi-tilt EMT data sets.

Indexed as

ChromatinElectron Microscope TomographyImaging, Three-DimensionalAnimalsCell NucleusDNAHumansNucleosomesChromatinDNANucleosomes

Identifiers

PMID39613943
PMCPMC12928153

What OpenQuestion holds

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Registered trials

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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.