---
DOAJ_listed: '1'
OA_place: publisher
OA_type: gold
PlanS_conform: '1'
_id: '20370'
abstract:
- lang: eng
  text: The Huntingtin protein (HTT), named for its role in Huntington’s disease,
    has been best understood as a scaffolding protein that promotes vesicle transport
    by molecular motors along microtubules. Here, we show that HTT also interacts
    with the actin cytoskeleton, and its loss of function disturbs the morphology
    and function of the axonal growth cone. We demonstrate that HTT organizes F-actin
    into bundles. Cryo–electron tomography (cryo-ET) and subtomogram averaging (STA)
    structural analyses reveal that HTT’s N-terminal HEAT and Bridge domains wrap
    around F-actin, while the C-terminal HEAT domain is displaced; furthermore, HTT
    dimerizes via the N-HEAT domain to bridge parallel actin filaments separated by
    ~20 nanometers. Our study provides the structural basis for understanding how
    HTT interacts with and organizes the actin cytoskeleton.
acknowledgement: 'We thank C. Cuveillier, J. Delaroche, T. Ferraro, and A. Zanchi
  for help with TIRF experiments, electron microscopy preparation, data analysis,
  and cell cultures, respectively; A. Antkowiak, C. Bosc, C. Fassier, A. Fourest-Lieuvin,
  and V. Brandt for helpful discussions. We acknowledge the contribution of the Photonic
  Imaging Center of Grenoble Institute Neuroscience which is part of the ISdV core
  facility and certified by the IBiSA label and ICM.Quant (RRID:SCR_026393) core facility
  of the Paris Brain Institute (ICM); the AniRA lentivector production facility from
  the CELPHEDIA Infrastructure and SFR Biosciences (UAR3444/CNRS, US8/Inserm, ENS
  de Lyon, UCBL); the Scientific Service Units (SSUs) of ISTA through resources provided
  by Scientific Computing (SciComp, A. Schloegl and S. Elefante); and the Electron
  Microscopy Facility (EMF, V.V. Hodirnau). The software programs used for the processing
  were supported by SBGrid (www.sbgrid.org). This work was supported by the Agence
  Nationale pour la Recherche (AXYON: ANR-18-CE16-0009-01, S.H.), Austrian Science
  Fund (FWF) grants (P33367, F.K.M.S.; E435, J.M.H.), ChanZuckerberg Initiative (CZI)
  grant (DAF2021-234754, F.K.M.S.), Hereditary Disease Foundation Research Grant (HDF
  990846, M.C.), European Union (ERC: ActinID 101076260, F.K.M.S.), Fondation pour
  la Recherche Médicale (FRM: équipe labellisée DEQ202203014675, S.H.; PhD fellowship,
  FDT202001010865, R.C.), Korea Health Industry Development Institute (KHIDI) (Korea-Switzerland
  global research support grant: RS-2023-00266300, J.-J.S.), National Research Foundation
  (NRF) of Korea (Korea-Austria collaborative grant NRF-2019K1A3A1A181160, J.-J.S.
  and F.K.M.S.; NRF-2020R1A2B5B03001517 and RS-2024-00333346 and RS-2024-00436173,
  J.-J.S.; 2021R1C1C1006700, D.K.).'
article_number: eadw4124
article_processing_charge: Yes
article_type: original
author:
- first_name: Rémi
  full_name: Carpentier, Rémi
  last_name: Carpentier
- first_name: Jaesung
  full_name: Kim, Jaesung
  last_name: Kim
- first_name: Mariacristina
  full_name: Capizzi, Mariacristina
  last_name: Capizzi
- first_name: Hyeongju
  full_name: Kim, Hyeongju
  last_name: Kim
- first_name: Florian
  full_name: Fäßler, Florian
  id: 404F5528-F248-11E8-B48F-1D18A9856A87
  last_name: Fäßler
  orcid: 0000-0001-7149-769X
- first_name: Jesse
  full_name: Hansen, Jesse
  id: 1063c618-6f9b-11ec-9123-f912fccded63
  last_name: Hansen
  orcid: 0000-0001-7967-2085
- first_name: Min Jeong
  full_name: Kim, Min Jeong
  last_name: Kim
- first_name: Eric
  full_name: Denarier, Eric
  last_name: Denarier
- first_name: Béatrice
  full_name: Blot, Béatrice
  last_name: Blot
- first_name: Marine
  full_name: Degennaro, Marine
  last_name: Degennaro
- first_name: Sophia
  full_name: Labou, Sophia
  last_name: Labou
- first_name: Isabelle
  full_name: Arnal, Isabelle
  last_name: Arnal
- first_name: Maria J.
  full_name: Marcaida, Maria J.
  last_name: Marcaida
- first_name: Matteo Dal
  full_name: Peraro, Matteo Dal
  last_name: Peraro
- first_name: Doory
  full_name: Kim, Doory
  last_name: Kim
- first_name: Florian KM
  full_name: Schur, Florian KM
  id: 48AD8942-F248-11E8-B48F-1D18A9856A87
  last_name: Schur
  orcid: 0000-0003-4790-8078
- first_name: Ji-Joon
  full_name: Song, Ji-Joon
  last_name: Song
- first_name: Sandrine
  full_name: Humbert, Sandrine
  last_name: Humbert
citation:
  ama: Carpentier R, Kim J, Capizzi M, et al. Structure of the Huntingtin F-actin
    complex reveals its role in cytoskeleton organization. <i>Science Advances</i>.
    2025;11(38). doi:<a href="https://doi.org/10.1126/sciadv.adw4124">10.1126/sciadv.adw4124</a>
  apa: Carpentier, R., Kim, J., Capizzi, M., Kim, H., Fäßler, F., Hansen, J., … Humbert,
    S. (2025). Structure of the Huntingtin F-actin complex reveals its role in cytoskeleton
    organization. <i>Science Advances</i>. AAAS. <a href="https://doi.org/10.1126/sciadv.adw4124">https://doi.org/10.1126/sciadv.adw4124</a>
  chicago: Carpentier, Rémi, Jaesung Kim, Mariacristina Capizzi, Hyeongju Kim, Florian
    Fäßler, Jesse Hansen, Min Jeong Kim, et al. “Structure of the Huntingtin F-Actin
    Complex Reveals Its Role in Cytoskeleton Organization.” <i>Science Advances</i>.
    AAAS, 2025. <a href="https://doi.org/10.1126/sciadv.adw4124">https://doi.org/10.1126/sciadv.adw4124</a>.
  ieee: R. Carpentier <i>et al.</i>, “Structure of the Huntingtin F-actin complex
    reveals its role in cytoskeleton organization,” <i>Science Advances</i>, vol.
    11, no. 38. AAAS, 2025.
  ista: Carpentier R, Kim J, Capizzi M, Kim H, Fäßler F, Hansen J, Kim MJ, Denarier
    E, Blot B, Degennaro M, Labou S, Arnal I, Marcaida MJ, Peraro MD, Kim D, Schur
    FK, Song J-J, Humbert S. 2025. Structure of the Huntingtin F-actin complex reveals
    its role in cytoskeleton organization. Science Advances. 11(38), eadw4124.
  mla: Carpentier, Rémi, et al. “Structure of the Huntingtin F-Actin Complex Reveals
    Its Role in Cytoskeleton Organization.” <i>Science Advances</i>, vol. 11, no.
    38, eadw4124, AAAS, 2025, doi:<a href="https://doi.org/10.1126/sciadv.adw4124">10.1126/sciadv.adw4124</a>.
  short: R. Carpentier, J. Kim, M. Capizzi, H. Kim, F. Fäßler, J. Hansen, M.J. Kim,
    E. Denarier, B. Blot, M. Degennaro, S. Labou, I. Arnal, M.J. Marcaida, M.D. Peraro,
    D. Kim, F.K. Schur, J.-J. Song, S. Humbert, Science Advances 11 (2025).
corr_author: '1'
date_created: 2025-09-22T08:00:52Z
date_published: 2025-09-19T00:00:00Z
date_updated: 2026-02-16T11:45:54Z
day: '19'
ddc:
- '570'
department:
- _id: FlSc
doi: 10.1126/sciadv.adw4124
external_id:
  isi:
  - '001575751700013'
  pmid:
  - '40971423'
file:
- access_level: open_access
  checksum: 4e2407bdabf8d53f399eb8a20d86218e
  content_type: application/pdf
  creator: dernst
  date_created: 2025-09-23T07:57:51Z
  date_updated: 2025-09-23T07:57:51Z
  file_id: '20372'
  file_name: 2025_ScienceAdvance_Carpentier.pdf
  file_size: 3599137
  relation: main_file
  success: 1
file_date_updated: 2025-09-23T07:57:51Z
has_accepted_license: '1'
intvolume: '        11'
isi: 1
issue: '38'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '09'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 9B954C5C-BA93-11EA-9121-9846C619BF3A
  grant_number: P33367
  name: Structure and isoform diversity of the Arp2/3 complex
- _id: 7bd318a1-9f16-11ee-852c-cc9217763180
  grant_number: E435
  name: In Situ Actin Structures via Hybrid Cryo-electron Microscopy
- _id: 62909c6f-2b32-11ec-9570-e1476aab5308
  grant_number: CZI01
  name: CryoMinflux-guided in-situ molecular census and structure determination
- _id: bd980d18-d553-11ed-ba76-ceaa645c97eb
  grant_number: '101076260'
  name: A molecular atlas of Actin filament IDentities in the cell motility machinery
publication: Science Advances
publication_identifier:
  issn:
  - 2375-2548
publication_status: published
publisher: AAAS
quality_controlled: '1'
scopus_import: '1'
status: public
title: Structure of the Huntingtin F-actin complex reveals its role in cytoskeleton
  organization
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 11
year: '2025'
...
---
DOAJ_listed: '1'
OA_place: publisher
OA_type: gold
_id: '19795'
abstract:
- lang: eng
  text: Super-resolution microscopy often entails long acquisition times of minutes
    to hours. Since drifts during the acquisition adversely affect data quality, active
    sample stabilization is commonly used for some of these techniques to reach their
    full potential. Although drifts in the lateral plane can often be corrected after
    acquisition, this is not always possible or may come with drawbacks. Therefore,
    it is appealing to stabilize sample position in three dimensions (3D) during acquisition.
    Various schemes for active sample stabilization have been demonstrated previously,
    with some reaching sub-nanometer stability in 3D. Here, we present a scheme for
    active drift correction that delivers the nanometer-scale 3D stability demanded
    by state-of-the-art super-resolution techniques and is straightforward to implement
    compared to previous schemes capable of reaching this level of stabilization precision.
    Using a refined algorithm that can handle various types of reference structure,
    without sparse signal peaks being mandatory, we stabilized sample position to
    ∼1 nm in 3D using objective lenses both with high and low numerical aperture.
    Our implementation requires only the addition of a simple widefield imaging path
    and we provide an open-source control software with graphical user interface to
    facilitate easy adoption of the module. Finally, we demonstrate how this has the
    potential to enhance data collection for diffraction-limited and super-resolution
    imaging techniques using single-molecule localization microscopy and cryo-confocal
    imaging as showcases.
acknowledged_ssus:
- _id: M-Shop
- _id: EM-Fac
- _id: LifeSc
acknowledgement: 'We acknowledge expert support by ISTA’s scientific service units,
  including the Miba Machine Shop, the Electron Microscopy Facility, and the Lab Support
  Facility. This work has been made possible in part by CZI grant DAF2021-234754 and
  grant DOI: https://doi.org/10.37921/812628ebpcwg from the Chan Zuckerberg Initiative
  DAF, an advised fund of Silicon Valley Community Foundation (funder DOI: https://doi.org/10.13039/100014989)
  (F.K.M.S. and J.G.D.). We further gratefully acknowledge funding by the following
  sources: Austrian Science Fund (FWF) grant DK W1232 (M.R.T. and J.G.D.); Austrian
  Academy of Sciences DOC fellowship 26137 (M.R.T.); Marie Skłodowska-Curie Actions
  Fellowship GA no. 665385 under the EU Horizon 2020 program (J.L.); ISTA postdoctoral
  fellowship IST fellow (A.W.); and Human Frontier Science Program postdoctoral fellowship
  LT000557/2018 (W.J.).'
article_number: '100211'
article_processing_charge: Yes
article_type: original
author:
- first_name: Jakob
  full_name: Vorlaufer, Jakob
  id: 937696FA-C996-11E9-8C7C-CF13E6697425
  last_name: Vorlaufer
  orcid: 0009-0000-7590-3501
- first_name: Nikolai
  full_name: Semenov, Nikolai
  id: e64d39c7-72ef-11ef-b75a-ee3046860d1b
  last_name: Semenov
- first_name: Caroline
  full_name: Kreuzinger, Caroline
  id: 382077BA-F248-11E8-B48F-1D18A9856A87
  last_name: Kreuzinger
- first_name: Manjunath
  full_name: Javoor, Manjunath
  id: 305ab18b-dc7d-11ea-9b2f-b58195228ea2
  last_name: Javoor
  orcid: 0000-0003-2311-2112
- first_name: Bettina
  full_name: Zens, Bettina
  id: 45FD126C-F248-11E8-B48F-1D18A9856A87
  last_name: Zens
  orcid: 0000-0002-9561-1239
- first_name: Nathalie
  full_name: Agudelo Duenas, Nathalie
  id: 40E7F008-F248-11E8-B48F-1D18A9856A87
  last_name: Agudelo Duenas
- first_name: Mojtaba
  full_name: Tavakoli, Mojtaba
  id: 3A0A06F4-F248-11E8-B48F-1D18A9856A87
  last_name: Tavakoli
  orcid: 0000-0002-7667-6854
- first_name: Marek
  full_name: Suplata, Marek
  id: EE8452B8-C26A-11E9-B157-E80CE6697425
  last_name: Suplata
- first_name: Wiebke
  full_name: Jahr, Wiebke
  id: 425C1CE8-F248-11E8-B48F-1D18A9856A87
  last_name: Jahr
  orcid: 0000-0003-0201-2315
- first_name: Julia
  full_name: Lyudchik, Julia
  id: 46E28B80-F248-11E8-B48F-1D18A9856A87
  last_name: Lyudchik
- first_name: Andreas
  full_name: Wartak, Andreas
  id: 60aaa06c-3de5-11eb-9e53-baa88e955dcb
  last_name: Wartak
- first_name: Florian Km
  full_name: Schur, Florian Km
  id: 48AD8942-F248-11E8-B48F-1D18A9856A87
  last_name: Schur
  orcid: 0000-0003-4790-8078
- first_name: Johann G
  full_name: Danzl, Johann G
  id: 42EFD3B6-F248-11E8-B48F-1D18A9856A87
  last_name: Danzl
  orcid: 0000-0001-8559-3973
citation:
  ama: Vorlaufer J, Semenov N, Kreuzinger C, et al. Image-based 3D active sample stabilization
    on the nanometer scale for optical microscopy. <i>Biophysical Reports</i>. 2025;5(2).
    doi:<a href="https://doi.org/10.1016/j.bpr.2025.100211">10.1016/j.bpr.2025.100211</a>
  apa: Vorlaufer, J., Semenov, N., Kreuzinger, C., Javoor, M., Zens, B., Agudelo Duenas,
    N., … Danzl, J. G. (2025). Image-based 3D active sample stabilization on the nanometer
    scale for optical microscopy. <i>Biophysical Reports</i>. Elsevier. <a href="https://doi.org/10.1016/j.bpr.2025.100211">https://doi.org/10.1016/j.bpr.2025.100211</a>
  chicago: Vorlaufer, Jakob, Nikolai Semenov, Caroline Kreuzinger, Manjunath Javoor,
    Bettina Zens, Nathalie Agudelo Duenas, Mojtaba Tavakoli, et al. “Image-Based 3D
    Active Sample Stabilization on the Nanometer Scale for Optical Microscopy.” <i>Biophysical
    Reports</i>. Elsevier, 2025. <a href="https://doi.org/10.1016/j.bpr.2025.100211">https://doi.org/10.1016/j.bpr.2025.100211</a>.
  ieee: J. Vorlaufer <i>et al.</i>, “Image-based 3D active sample stabilization on
    the nanometer scale for optical microscopy,” <i>Biophysical Reports</i>, vol.
    5, no. 2. Elsevier, 2025.
  ista: Vorlaufer J, Semenov N, Kreuzinger C, Javoor M, Zens B, Agudelo Duenas N,
    Tavakoli M, Suplata M, Jahr W, Lyudchik J, Wartak A, Schur FK, Danzl JG. 2025.
    Image-based 3D active sample stabilization on the nanometer scale for optical
    microscopy. Biophysical Reports. 5(2), 100211.
  mla: Vorlaufer, Jakob, et al. “Image-Based 3D Active Sample Stabilization on the
    Nanometer Scale for Optical Microscopy.” <i>Biophysical Reports</i>, vol. 5, no.
    2, 100211, Elsevier, 2025, doi:<a href="https://doi.org/10.1016/j.bpr.2025.100211">10.1016/j.bpr.2025.100211</a>.
  short: J. Vorlaufer, N. Semenov, C. Kreuzinger, M. Javoor, B. Zens, N. Agudelo Duenas,
    M. Tavakoli, M. Suplata, W. Jahr, J. Lyudchik, A. Wartak, F.K. Schur, J.G. Danzl,
    Biophysical Reports 5 (2025).
corr_author: '1'
date_created: 2025-06-08T22:01:22Z
date_published: 2025-06-11T00:00:00Z
date_updated: 2026-04-07T11:48:07Z
day: '11'
ddc:
- '570'
department:
- _id: JoDa
- _id: GradSch
- _id: FlSc
- _id: EM-Fac
doi: 10.1016/j.bpr.2025.100211
ec_funded: 1
file:
- access_level: open_access
  checksum: 4018c833f25a3ad3b57e3577fed70334
  content_type: application/pdf
  creator: dernst
  date_created: 2025-06-10T07:24:46Z
  date_updated: 2025-06-10T07:24:46Z
  file_id: '19802'
  file_name: 2025_BiophysicalReports_Vorlaufer.pdf
  file_size: 7238179
  relation: main_file
  success: 1
file_date_updated: 2025-06-10T07:24:46Z
has_accepted_license: '1'
intvolume: '         5'
issue: '2'
language:
- iso: eng
month: '06'
oa: 1
oa_version: Published Version
project:
- _id: 62909c6f-2b32-11ec-9570-e1476aab5308
  grant_number: CZI01
  name: CryoMinflux-guided in-situ molecular census and structure determination
- _id: 6285a163-2b32-11ec-9570-8e204ca2dba5
  grant_number: '26137'
  name: Studying Organelle Structure and Function at Nanoscale Resolution with Expansion
    Microscopy
- _id: 2564DBCA-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '665385'
  name: International IST Doctoral Program
- _id: 26AA4EF2-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: W1232-B24
  name: Molecular Drug Targets
- _id: 2668BFA0-B435-11E9-9278-68D0E5697425
  grant_number: LT00057
  name: High-speed 3D-nanoscopy to study the role of adhesion during 3D cell migration
publication: Biophysical Reports
publication_identifier:
  eissn:
  - 2667-0747
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  record:
  - id: '20206'
    relation: dissertation_contains
    status: public
scopus_import: '1'
status: public
title: Image-based 3D active sample stabilization on the nanometer scale for optical
  microscopy
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 5
year: '2025'
...
---
OA_place: publisher
_id: '20206'
abstract:
- lang: eng
  text: "The internal structure of biomolecules and their organization in higher-order
    arrangements are key factors governing the working principles of biological systems.
    Bioimaging has successfully revealed arrangements across relevant spatial scales.
    For example, cryo-electron tomography has become widely used for analyzing biomolecular
    structures in situ due to its comprehensive structural visualization of near-natively
    preserved samples, and its capability of sub-nm resolution via averaging. However,
    the identification of molecules within crowded cellular environments is often
    hindered by low contrast. Fluorescence microscopy, on the other hand, routinely
    visualizes specifically labeled targets at single-molecule contrast against essentially
    zero background. Moreover, it provides comparatively high throughput and is amenable
    to multiplexing. Due to this complementarity, combining datasets from both modalities
    acquired on the same region via correlative light and electron microscopy can
    reveal novel types of information. \r\nThe spatial scale at which information
    can be extracted depends on imaging resolution and correlation accuracy. Since
    diffraction of light limits the resolution of conventional fluorescence microscopy
    to few hundreds of nanometers, reaching the full potential of correlative imaging
    requires super-resolution approaches. Performing imaging at cryogenic temperature
    preserves structures in a near-native state and minimizes distortions between
    the fluorescence and the electron microscopy datasets. Implementations of this
    concept have achieved correlation on the scale of cellular organelles or bacterial
    domains.\r\nWe have worked towards pushing correlative imaging to the single-molecule
    scale by improving cryo-super-resolution microscopy, and devising a refined image
    correlation workflow. As part of this project, I constructed a microscopy setup
    and adopted it for super-resolution fluorescence microscopy at room temperature
    and cryogenic conditions. I explored different cryo-stages and acquisition strategies.
    Specifically, I developed a new scheme for correcting sample drift, thus increasing
    mechanical stability during microscopy acquisitions.\r\n"
acknowledged_ssus:
- _id: M-Shop
- _id: EM-Fac
- _id: Bio
acknowledgement: "The project was supported by CZI grant DAF2021-234754 and grant\r\nDOI:
  https://doi.org/10.37921/812628ebpcwg from the Chan Zuckerberg Initiative DAF, an\r\nadvised
  fund of Silicon Valley Community Foundation (funder\r\nDOI: https://doi.org/10.13039/100014989),
  as well as internal grants from ISTA’s Equipment\r\nInvestment Committee and Interdisciplinary
  Project Committee. "
alternative_title:
- ISTA Thesis
article_processing_charge: No
author:
- first_name: Jakob
  full_name: Vorlaufer, Jakob
  id: 937696FA-C996-11E9-8C7C-CF13E6697425
  last_name: Vorlaufer
  orcid: 0009-0000-7590-3501
citation:
  ama: Vorlaufer J. Construction of a cryo-super-resolution microscope to guide in
    situ structure analysis. 2025. doi:<a href="https://doi.org/10.15479/AT-ISTA-20206">10.15479/AT-ISTA-20206</a>
  apa: Vorlaufer, J. (2025). <i>Construction of a cryo-super-resolution microscope
    to guide in situ structure analysis</i>. Institute of Science and Technology Austria.
    <a href="https://doi.org/10.15479/AT-ISTA-20206">https://doi.org/10.15479/AT-ISTA-20206</a>
  chicago: Vorlaufer, Jakob. “Construction of a Cryo-Super-Resolution Microscope to
    Guide in Situ Structure Analysis.” Institute of Science and Technology Austria,
    2025. <a href="https://doi.org/10.15479/AT-ISTA-20206">https://doi.org/10.15479/AT-ISTA-20206</a>.
  ieee: J. Vorlaufer, “Construction of a cryo-super-resolution microscope to guide
    in situ structure analysis,” Institute of Science and Technology Austria, 2025.
  ista: Vorlaufer J. 2025. Construction of a cryo-super-resolution microscope to guide
    in situ structure analysis. Institute of Science and Technology Austria.
  mla: Vorlaufer, Jakob. <i>Construction of a Cryo-Super-Resolution Microscope to
    Guide in Situ Structure Analysis</i>. Institute of Science and Technology Austria,
    2025, doi:<a href="https://doi.org/10.15479/AT-ISTA-20206">10.15479/AT-ISTA-20206</a>.
  short: J. Vorlaufer, Construction of a Cryo-Super-Resolution Microscope to Guide
    in Situ Structure Analysis, Institute of Science and Technology Austria, 2025.
corr_author: '1'
date_created: 2025-08-22T08:12:55Z
date_published: 2025-08-25T00:00:00Z
date_updated: 2026-04-07T11:48:07Z
day: '25'
ddc:
- '621'
- '535'
degree_awarded: PhD
department:
- _id: GradSch
- _id: JoDa
doi: 10.15479/AT-ISTA-20206
file:
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language:
- iso: eng
license: https://creativecommons.org/licenses/by-nc-sa/4.0/
month: '08'
oa: 1
oa_version: Published Version
page: '107'
project:
- _id: 62909c6f-2b32-11ec-9570-e1476aab5308
  grant_number: CZI01
  name: CryoMinflux-guided in-situ molecular census and structure determination
publication_identifier:
  issn:
  - 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
  record:
  - id: '19795'
    relation: part_of_dissertation
    status: public
status: public
supervisor:
- first_name: Johann G
  full_name: Danzl, Johann G
  id: 42EFD3B6-F248-11E8-B48F-1D18A9856A87
  last_name: Danzl
  orcid: 0000-0001-8559-3973
title: Construction of a cryo-super-resolution microscope to guide in situ structure
  analysis
tmp:
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  name: Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC
    BY-NC-SA 4.0)
  short: CC BY-NC-SA (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2025'
...
---
_id: '15146'
abstract:
- lang: eng
  text: The extracellular matrix (ECM) serves as a scaffold for cells and plays an
    essential role in regulating numerous cellular processes, including cell migration
    and proliferation. Due to limitations in specimen preparation for conventional
    room-temperature electron microscopy, we lack structural knowledge on how ECM
    components are secreted, remodeled, and interact with surrounding cells. We have
    developed a 3D-ECM platform compatible with sample thinning by cryo-focused ion
    beam milling, the lift-out extraction procedure, and cryo-electron tomography.
    Our workflow implements cell-derived matrices (CDMs) grown on EM grids, resulting
    in a versatile tool closely mimicking ECM environments. This allows us to visualize
    ECM for the first time in its hydrated, native context. Our data reveal an intricate
    network of extracellular fibers, their positioning relative to matrix-secreting
    cells, and previously unresolved structural entities. Our workflow and results
    add to the structural atlas of the ECM, providing novel insights into its secretion
    and assembly.
acknowledged_ssus:
- _id: LifeSc
- _id: ScienComp
- _id: EM-Fac
- _id: M-Shop
acknowledgement: "Open Access funding provided by IST Austria. We thank Armel Nicolas
  and his team at the ISTA proteomics facility, Alois Schloegl, Stefano Elefante,
  and colleagues at the ISTA Scientific Computing facility, Tommaso Constanzo and
  Ludek Lovicar at the Electron Microsocpy Facility (EMF), and Thomas Menner at the
  Miba Machine shop for their support. We also thank Wanda Kukulski (University of
  Bern) as well as Darío Porley, Andreas Thader, and other members of the Schur group
  for helpful discussions. Matt Swulius and Jessica Heebner provided great support
  in using Dragonfly. We thank Dorotea Fracciolla (Art & Science) for support in figure
  illustration.\r\n\r\nThis research was supported by the Scientific Service Units
  of ISTA through resources provided by Scientific Computing, the Lab Support Facility,
  and the Electron Microscopy Facility. We acknowledge funding support from the following
  sources: Austrian Science Fund (FWF) grant P33367 (to F.K.M. Schur), the Federation
  of European Biochemical Societies (to F.K.M. Schur), Niederösterreich (NÖ) Fonds
  (to B. Zens), FWF grant E435 (to J.M. Hansen), European Research Council under the
  European Union’s Horizon 2020 research (grant agreement No. 724373) (to M. Sixt),
  and Jenny and Antti Wihuri Foundation (to J. Alanko). This publication has been
  made possible in part by CZI grant DAF2021-234754 and grant DOI https://doi.org/10.37921/812628ebpcwg
  from the Chan Zuckerberg Initiative DAF, an advised fund of Silicon Valley Community
  Foundation (to F.K.M. Schur)."
article_number: e202309125
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Bettina
  full_name: Zens, Bettina
  id: 45FD126C-F248-11E8-B48F-1D18A9856A87
  last_name: Zens
  orcid: 0000-0002-9561-1239
- first_name: Florian
  full_name: Fäßler, Florian
  id: 404F5528-F248-11E8-B48F-1D18A9856A87
  last_name: Fäßler
  orcid: 0000-0001-7149-769X
- first_name: Jesse
  full_name: Hansen, Jesse
  id: 1063c618-6f9b-11ec-9123-f912fccded63
  last_name: Hansen
  orcid: 0000-0001-7967-2085
- first_name: Robert
  full_name: Hauschild, Robert
  id: 4E01D6B4-F248-11E8-B48F-1D18A9856A87
  last_name: Hauschild
  orcid: 0000-0001-9843-3522
- first_name: Julia
  full_name: Datler, Julia
  id: 3B12E2E6-F248-11E8-B48F-1D18A9856A87
  last_name: Datler
  orcid: 0000-0002-3616-8580
- first_name: Victor-Valentin
  full_name: Hodirnau, Victor-Valentin
  id: 3661B498-F248-11E8-B48F-1D18A9856A87
  last_name: Hodirnau
  orcid: 0000-0003-3904-947X
- first_name: Vanessa
  full_name: Zheden, Vanessa
  id: 39C5A68A-F248-11E8-B48F-1D18A9856A87
  last_name: Zheden
  orcid: 0000-0002-9438-4783
- first_name: Jonna H
  full_name: Alanko, Jonna H
  id: 2CC12E8C-F248-11E8-B48F-1D18A9856A87
  last_name: Alanko
  orcid: 0000-0002-7698-3061
- first_name: Michael K
  full_name: Sixt, Michael K
  id: 41E9FBEA-F248-11E8-B48F-1D18A9856A87
  last_name: Sixt
  orcid: 0000-0002-6620-9179
- first_name: Florian KM
  full_name: Schur, Florian KM
  id: 48AD8942-F248-11E8-B48F-1D18A9856A87
  last_name: Schur
  orcid: 0000-0003-4790-8078
citation:
  ama: Zens B, Fäßler F, Hansen J, et al. Lift-out cryo-FIBSEM and cryo-ET reveal
    the ultrastructural landscape of extracellular matrix. <i>Journal of Cell Biology</i>.
    2024;223(6). doi:<a href="https://doi.org/10.1083/jcb.202309125">10.1083/jcb.202309125</a>
  apa: Zens, B., Fäßler, F., Hansen, J., Hauschild, R., Datler, J., Hodirnau, V.-V.,
    … Schur, F. K. (2024). Lift-out cryo-FIBSEM and cryo-ET reveal the ultrastructural
    landscape of extracellular matrix. <i>Journal of Cell Biology</i>. Rockefeller
    University Press. <a href="https://doi.org/10.1083/jcb.202309125">https://doi.org/10.1083/jcb.202309125</a>
  chicago: Zens, Bettina, Florian Fäßler, Jesse Hansen, Robert Hauschild, Julia Datler,
    Victor-Valentin Hodirnau, Vanessa Zheden, Jonna H Alanko, Michael K Sixt, and
    Florian KM Schur. “Lift-out Cryo-FIBSEM and Cryo-ET Reveal the Ultrastructural
    Landscape of Extracellular Matrix.” <i>Journal of Cell Biology</i>. Rockefeller
    University Press, 2024. <a href="https://doi.org/10.1083/jcb.202309125">https://doi.org/10.1083/jcb.202309125</a>.
  ieee: B. Zens <i>et al.</i>, “Lift-out cryo-FIBSEM and cryo-ET reveal the ultrastructural
    landscape of extracellular matrix,” <i>Journal of Cell Biology</i>, vol. 223,
    no. 6. Rockefeller University Press, 2024.
  ista: Zens B, Fäßler F, Hansen J, Hauschild R, Datler J, Hodirnau V-V, Zheden V,
    Alanko JH, Sixt MK, Schur FK. 2024. Lift-out cryo-FIBSEM and cryo-ET reveal the
    ultrastructural landscape of extracellular matrix. Journal of Cell Biology. 223(6),
    e202309125.
  mla: Zens, Bettina, et al. “Lift-out Cryo-FIBSEM and Cryo-ET Reveal the Ultrastructural
    Landscape of Extracellular Matrix.” <i>Journal of Cell Biology</i>, vol. 223,
    no. 6, e202309125, Rockefeller University Press, 2024, doi:<a href="https://doi.org/10.1083/jcb.202309125">10.1083/jcb.202309125</a>.
  short: B. Zens, F. Fäßler, J. Hansen, R. Hauschild, J. Datler, V.-V. Hodirnau, V.
    Zheden, J.H. Alanko, M.K. Sixt, F.K. Schur, Journal of Cell Biology 223 (2024).
corr_author: '1'
date_created: 2024-03-21T06:45:51Z
date_published: 2024-03-20T00:00:00Z
date_updated: 2025-09-04T13:17:16Z
day: '20'
ddc:
- '570'
department:
- _id: FlSc
- _id: MiSi
- _id: Bio
- _id: EM-Fac
doi: 10.1083/jcb.202309125
ec_funded: 1
external_id:
  isi:
  - '001264190100001'
  pmid:
  - '38506714'
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  creator: dernst
  date_created: 2024-03-25T12:52:04Z
  date_updated: 2024-03-25T12:52:04Z
  file_id: '15188'
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file_date_updated: 2024-03-25T12:52:04Z
has_accepted_license: '1'
intvolume: '       223'
isi: 1
issue: '6'
language:
- iso: eng
month: '03'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 9B954C5C-BA93-11EA-9121-9846C619BF3A
  grant_number: P33367
  name: Structure and isoform diversity of the Arp2/3 complex
- _id: 7bd318a1-9f16-11ee-852c-cc9217763180
  grant_number: E435
  name: In Situ Actin Structures via Hybrid Cryo-electron Microscopy
- _id: 25FE9508-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '724373'
  name: Cellular Navigation Along Spatial Gradients
- _id: 059B463C-7A3F-11EA-A408-12923DDC885E
  name: "NÃ\x96-Fonds Preis für die Jungforscherin des Jahres am IST Austria"
- _id: 2615199A-B435-11E9-9278-68D0E5697425
  grant_number: '21317'
  name: Spatiotemporal regulation of chemokine-induced signalling in leukocyte chemotaxis
- _id: 62909c6f-2b32-11ec-9570-e1476aab5308
  grant_number: CZI01
  name: CryoMinflux-guided in-situ visual proteomics and structure determination
publication: Journal of Cell Biology
publication_identifier:
  eissn:
  - 1540-8140
  issn:
  - 0021-9525
publication_status: published
publisher: Rockefeller University Press
quality_controlled: '1'
scopus_import: '1'
status: public
title: Lift-out cryo-FIBSEM and cryo-ET reveal the ultrastructural landscape of extracellular
  matrix
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 223
year: '2024'
...