Evidence map›Paper›PMID 40773232›Full record

ArticleProceedings of the National Academy of Sciences of the United States of America2025

Cryosectioning-enhanced super-resolution microscopy for single-protein imaging across cells and tissues.

Johannes Stein, Maria Ericsson, Michel Nofal, Lorenzo Magni, Sarah Aufmkolk, Ryan B McMillan, Laura Breimann, Conor P Herlihy, S Dean Lee, Andréa Willemin and 6 more

Abstract read
In one paragraph

Article in Proceedings of the National Academy of Sciences of the United States of America, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing 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

4 citing papers in PubMed.

  1. Article
  2. Article
  3. Article
  4. Review
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

16 authors.

Johannes SteinWyss Institute of Biologically Inspired Engineering, Boston, MA 02115.ORCID 0000-0002-1335-1120
Maria EricssonBlavatnik Institute, Harvard Medical School, Boston, MA 02115.ORCID 0000-0002-4563-3683
Michel NofalWyss Institute of Biologically Inspired Engineering, Boston, MA 02115.ORCID 0000-0002-0994-6914
Lorenzo MagniWyss Institute of Biologically Inspired Engineering, Boston, MA 02115.
Sarah AufmkolkDepartment of Genetics, Harvard Medical School, Boston, MA 02115.
Ryan B McMillanWyss Institute of Biologically Inspired Engineering, Boston, MA 02115.ORCID 0000-0002-8905-0471
Laura BreimannDepartment of Genetics, Harvard Medical School, Boston, MA 02115.ORCID 0000-0001-7702-5749
Conor P HerlihyDepartment of Genetics, Harvard Medical School, Boston, MA 02115.ORCID 0000-0001-9818-4204
S Dean LeeDepartment of Genetics, Harvard Medical School, Boston, MA 02115.ORCID 0000-0001-6391-9821
Andréa WilleminMax-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin Institute for Medical Systems Biology, Epigenetic Regulation and Chromatin Architecture Group, Berlin 10115, Germany.
Jens WohlmannDepartment of Biosciences, University of Oslo, Oslo 0316, Norway.ORCID 0000-0001-7726-1557
Laura Arguedas-JimenezMax-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin Institute for Medical Systems Biology, Epigenetic Regulation and Chromatin Architecture Group, Berlin 10115, Germany.ORCID 0009-0004-2240-7781
Peng YinWyss Institute of Biologically Inspired Engineering, Boston, MA 02115.
Ana PomboMax-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin Institute for Medical Systems Biology, Epigenetic Regulation and Chromatin Architecture Group, Berlin 10115, Germany.ORCID 0000-0002-7493-6288
George M ChurchWyss Institute of Biologically Inspired Engineering, Boston, MA 02115.ORCID 0000-0003-3535-2076
Chao-Ting WuDepartment of Genetics, Harvard Medical School, Boston, MA 02115.ORCID 0000-0001-5525-871X

Funding

MOLECULAR BIOPHYSICS TRAINING GRANTT32GM008313 · NIGMS · HARVARD UNIVERSITY (MEDICAL SCHOOL) · PI MURTHY, VENKATESH N · 1989 to 2023
$14.6M
Center for Genome ImagingRM1HG011016 · NHGRI · HARVARD MEDICAL SCHOOL · PI WU, CHAO-TING · 2021 to 2025
$12.2M
Multiscale Analyses of 4D Nucleome Structure and Function by Comprehensive Multimodal Data IntegrationUM1HG011593 · NHGRI · CARNEGIE-MELLON UNIVERSITY · PI ALBER, FRANK, BELMONT, ANDREW STEVEN · 2020 to 2024
$10.4M
High-throughput single-molecule protein identification via super-resolution imagingDP1GM133052 · NIGMS · HARVARD UNIVERSITY · PI YIN, PENG · 2018 to 2022
$6.6M
European Molecular Biology Organization (EMBO) ALTF 816-2021HHS | NIH (NIH) 5RM1HG011016-03HHS | NIH (NIH) 5UM1HG011593HHS | NIH (NIH) DP1GM133052HHS | NIH (NIH) NIGMS T32 GM008313NHGRI NIH HHS RM1 HG011016NHGRI NIH HHS UM1 HG011593NIGMS NIH HHS DP1 GM133052NIGMS NIH HHS T32 GM008313NSF (NSF) 2140743U.S. Department of Energy (DOE) DE-FG02-02ER63445
6 · The paper itself

Abstract

DNA-points accumulation for imaging in nanoscale topography (DNA-PAINT) enables nanoscale imaging with virtually unlimited multiplexing and molecular counting. Here, we address challenges, such as variable imaging performance and target accessibility, that can limit its broader applicability. Specifically, we enhance its capacity for robust single-protein imaging and molecular counting by optimizing the integration of total internal reflection fluorescence microscopy with physical sectioning, in particular, Tokuyasu cryosectioning. Our method, tomographic and kinetically enhanced DNA-PAINT (tkPAINT), achieves 3 nm localization precision across diverse samples, enhanced imager binding, and improved cellular integrity. tkPAINT can facilitate molecular counting with DNA-PAINT inside the nucleus, as demonstrated through its quantification of the in situ abundance of RNA Polymerase II in both HeLa cells as well as mouse tissues. Anticipating that tkPAINT could become a versatile tool for the exploration of biomolecular organization and interactions across cells and tissues, we also demonstrate its capacity to support multiplexing, multimodal targeting of proteins and nucleic acids, and three-dimensional (3D) imaging.

Indexed as

CryoultramicrotomyProteinsSingle Molecule ImagingAnimalsDNAHeLa CellsHumansImaging, Three-DimensionalMiceMicroscopy, FluorescenceRNA Polymerase IIDNAProteinsRNA Polymerase IIDNA-PAINTnuclear organizationsingle-molecule localization microscopysuper-resolution microscopyTIRF

Identifiers

PMID40773232
PMCPMC12358845

What OpenQuestion holds

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LicenceCC BY-NC-ND
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Registered trials

None linked

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.