---
_id: '11155'
abstract:
- lang: eng
  text: The potential of energy filtering and direct electron detection for cryo-electron
    microscopy (cryo-EM) has been well documented. Here, we assess the performance
    of recently introduced hardware for cryo-electron tomography (cryo-ET) and subtomogram
    averaging (STA), an increasingly popular structural determination method for complex
    3D specimens. We acquired cryo-ET datasets of EIAV virus-like particles (VLPs)
    on two contemporary cryo-EM systems equipped with different energy filters and
    direct electron detectors (DED), specifically a Krios G4, equipped with a cold
    field emission gun (CFEG), Thermo Fisher Scientific Selectris X energy filter,
    and a Falcon 4 DED; and a Krios G3i, with a Schottky field emission gun (XFEG),
    a Gatan Bioquantum energy filter, and a K3 DED. We performed constrained cross-correlation-based
    STA on equally sized datasets acquired on the respective systems. The resulting
    EIAV CA hexamer reconstructions show that both systems perform comparably in the
    4–6 Å resolution range based on Fourier-Shell correlation (FSC). In addition,
    by employing a recently introduced multiparticle refinement approach, we obtained
    a reconstruction of the EIAV CA hexamer at 2.9 Å. Our results demonstrate the
    potential of the new generation of energy filters and DEDs for STA, and the effects
    of using different processing pipelines on their STA outcomes.
acknowledged_ssus:
- _id: LifeSc
- _id: ScienComp
- _id: EM-Fac
acknowledgement: This work was funded by the Austrian Science Fund (FWF) grant P31445
  to F.K.M.S and the National Institute of Allergy and Infectious Diseases under awards
  R01AI147890 to R.A.D. This research was also supported by the Scientific Service
  Units (SSUs) of IST Austria through resources provided by Scientific Computing (SciComp),
  the Life Science Facility (LSF), and the Electron Microscopy Facility (EMF). We
  thank Dustin Morado for providing the software SubTOM for data processing. We also
  thank William Wan for critical reading of the manuscript and valuable feedback.
article_number: '107852'
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Martin
  full_name: Obr, Martin
  id: 4741CA5A-F248-11E8-B48F-1D18A9856A87
  last_name: Obr
- first_name: Wim J.H.
  full_name: Hagen, Wim J.H.
  last_name: Hagen
- first_name: Robert A.
  full_name: Dick, Robert A.
  last_name: Dick
- first_name: Lingbo
  full_name: Yu, Lingbo
  last_name: Yu
- first_name: Abhay
  full_name: Kotecha, Abhay
  last_name: Kotecha
- 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: Obr M, Hagen WJH, Dick RA, Yu L, Kotecha A, Schur FK. Exploring high-resolution
    cryo-ET and subtomogram averaging capabilities of contemporary DEDs. <i>Journal
    of Structural Biology</i>. 2022;214(2). doi:<a href="https://doi.org/10.1016/j.jsb.2022.107852">10.1016/j.jsb.2022.107852</a>
  apa: Obr, M., Hagen, W. J. H., Dick, R. A., Yu, L., Kotecha, A., &#38; Schur, F.
    K. (2022). Exploring high-resolution cryo-ET and subtomogram averaging capabilities
    of contemporary DEDs. <i>Journal of Structural Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jsb.2022.107852">https://doi.org/10.1016/j.jsb.2022.107852</a>
  chicago: Obr, Martin, Wim J.H. Hagen, Robert A. Dick, Lingbo Yu, Abhay Kotecha,
    and Florian KM Schur. “Exploring High-Resolution Cryo-ET and Subtomogram Averaging
    Capabilities of Contemporary DEDs.” <i>Journal of Structural Biology</i>. Elsevier,
    2022. <a href="https://doi.org/10.1016/j.jsb.2022.107852">https://doi.org/10.1016/j.jsb.2022.107852</a>.
  ieee: M. Obr, W. J. H. Hagen, R. A. Dick, L. Yu, A. Kotecha, and F. K. Schur, “Exploring
    high-resolution cryo-ET and subtomogram averaging capabilities of contemporary
    DEDs,” <i>Journal of Structural Biology</i>, vol. 214, no. 2. Elsevier, 2022.
  ista: Obr M, Hagen WJH, Dick RA, Yu L, Kotecha A, Schur FK. 2022. Exploring high-resolution
    cryo-ET and subtomogram averaging capabilities of contemporary DEDs. Journal of
    Structural Biology. 214(2), 107852.
  mla: Obr, Martin, et al. “Exploring High-Resolution Cryo-ET and Subtomogram Averaging
    Capabilities of Contemporary DEDs.” <i>Journal of Structural Biology</i>, vol.
    214, no. 2, 107852, Elsevier, 2022, doi:<a href="https://doi.org/10.1016/j.jsb.2022.107852">10.1016/j.jsb.2022.107852</a>.
  short: M. Obr, W.J.H. Hagen, R.A. Dick, L. Yu, A. Kotecha, F.K. Schur, Journal of
    Structural Biology 214 (2022).
date_created: 2022-04-15T07:10:26Z
date_published: 2022-06-01T00:00:00Z
date_updated: 2023-08-03T06:25:23Z
day: '01'
ddc:
- '570'
department:
- _id: FlSc
doi: 10.1016/j.jsb.2022.107852
external_id:
  isi:
  - '000790733600001'
  pmid:
  - '35351542'
file:
- access_level: open_access
  checksum: 0b1eb53447aae8e95ae4c12d193b0b00
  content_type: application/pdf
  creator: dernst
  date_created: 2022-08-02T11:07:58Z
  date_updated: 2022-08-02T11:07:58Z
  file_id: '11722'
  file_name: 2022_JourStructuralBiology_Obr.pdf
  file_size: 7080863
  relation: main_file
  success: 1
file_date_updated: 2022-08-02T11:07:58Z
has_accepted_license: '1'
intvolume: '       214'
isi: 1
issue: '2'
keyword:
- Structural Biology
language:
- iso: eng
month: '06'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 26736D6A-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: P31445
  name: Structural conservation and diversity in retroviral capsid
publication: Journal of Structural Biology
publication_identifier:
  issn:
  - 1047-8477
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Exploring high-resolution cryo-ET and subtomogram averaging capabilities of
  contemporary DEDs
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: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 214
year: '2022'
...
---
_id: '10290'
abstract:
- lang: eng
  text: A precise quantitative description of the ultrastructural characteristics
    underlying biological mechanisms is often key to their understanding. This is
    particularly true for dynamic extra- and intracellular filamentous assemblies,
    playing a role in cell motility, cell integrity, cytokinesis, tissue formation
    and maintenance. For example, genetic manipulation or modulation of actin regulatory
    proteins frequently manifests in changes of the morphology, dynamics, and ultrastructural
    architecture of actin filament-rich cell peripheral structures, such as lamellipodia
    or filopodia. However, the observed ultrastructural effects often remain subtle
    and require sufficiently large datasets for appropriate quantitative analysis.
    The acquisition of such large datasets has been enabled by recent advances in
    high-throughput cryo-electron tomography (cryo-ET) methods. This also necessitates
    the development of complementary approaches to maximize the extraction of relevant
    biological information. We have developed a computational toolbox for the semi-automatic
    quantification of segmented and vectorized filamentous networks from pre-processed
    cryo-electron tomograms, facilitating the analysis and cross-comparison of multiple
    experimental conditions. GUI-based components simplify the processing of data
    and allow users to obtain a large number of ultrastructural parameters describing
    filamentous assemblies. We demonstrate the feasibility of this workflow by analyzing
    cryo-ET data of untreated and chemically perturbed branched actin filament networks
    and that of parallel actin filament arrays. In principle, the computational toolbox
    presented here is applicable for data analysis comprising any type of filaments
    in regular (i.e. parallel) or random arrangement. We show that it can ease the
    identification of key differences between experimental groups and facilitate the
    in-depth analysis of ultrastructural data in a time-efficient manner.
acknowledged_ssus:
- _id: ScienComp
- _id: LifeSc
- _id: Bio
- _id: EM-Fac
acknowledgement: 'This research was supported by the Scientific Service Units (SSUs)
  of IST Austria through resources provided by Scientific Computing (SciComp), the
  Life Science Facility (LSF), the BioImaging Facility (BIF), and the Electron Microscopy
  Facility (EMF). We also thank Victor-Valentin Hodirnau for help with cryo-ET data
  acquisition. The authors acknowledge support from IST Austria and from the Austrian
  Science Fund (FWF): M02495 to G.D. and Austrian Science Fund (FWF): P33367 to F.K.M.S.'
article_number: '107808'
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Georgi A
  full_name: Dimchev, Georgi A
  id: 38C393BE-F248-11E8-B48F-1D18A9856A87
  last_name: Dimchev
  orcid: 0000-0001-8370-6161
- first_name: Behnam
  full_name: Amiri, Behnam
  last_name: Amiri
- first_name: Florian
  full_name: Fäßler, Florian
  id: 404F5528-F248-11E8-B48F-1D18A9856A87
  last_name: Fäßler
  orcid: 0000-0001-7149-769X
- first_name: Martin
  full_name: Falcke, Martin
  last_name: Falcke
- 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: Dimchev GA, Amiri B, Fäßler F, Falcke M, Schur FK. Computational toolbox for
    ultrastructural quantitative analysis of filament networks in cryo-ET data. <i>Journal
    of Structural Biology</i>. 2021;213(4). doi:<a href="https://doi.org/10.1016/j.jsb.2021.107808">10.1016/j.jsb.2021.107808</a>
  apa: Dimchev, G. A., Amiri, B., Fäßler, F., Falcke, M., &#38; Schur, F. K. (2021).
    Computational toolbox for ultrastructural quantitative analysis of filament networks
    in cryo-ET data. <i>Journal of Structural Biology</i>. Elsevier . <a href="https://doi.org/10.1016/j.jsb.2021.107808">https://doi.org/10.1016/j.jsb.2021.107808</a>
  chicago: Dimchev, Georgi A, Behnam Amiri, Florian Fäßler, Martin Falcke, and Florian
    KM Schur. “Computational Toolbox for Ultrastructural Quantitative Analysis of
    Filament Networks in Cryo-ET Data.” <i>Journal of Structural Biology</i>. Elsevier
    , 2021. <a href="https://doi.org/10.1016/j.jsb.2021.107808">https://doi.org/10.1016/j.jsb.2021.107808</a>.
  ieee: G. A. Dimchev, B. Amiri, F. Fäßler, M. Falcke, and F. K. Schur, “Computational
    toolbox for ultrastructural quantitative analysis of filament networks in cryo-ET
    data,” <i>Journal of Structural Biology</i>, vol. 213, no. 4. Elsevier , 2021.
  ista: Dimchev GA, Amiri B, Fäßler F, Falcke M, Schur FK. 2021. Computational toolbox
    for ultrastructural quantitative analysis of filament networks in cryo-ET data.
    Journal of Structural Biology. 213(4), 107808.
  mla: Dimchev, Georgi A., et al. “Computational Toolbox for Ultrastructural Quantitative
    Analysis of Filament Networks in Cryo-ET Data.” <i>Journal of Structural Biology</i>,
    vol. 213, no. 4, 107808, Elsevier , 2021, doi:<a href="https://doi.org/10.1016/j.jsb.2021.107808">10.1016/j.jsb.2021.107808</a>.
  short: G.A. Dimchev, B. Amiri, F. Fäßler, M. Falcke, F.K. Schur, Journal of Structural
    Biology 213 (2021).
date_created: 2021-11-15T12:21:42Z
date_published: 2021-11-03T00:00:00Z
date_updated: 2023-11-21T08:36:02Z
day: '03'
ddc:
- '572'
department:
- _id: FlSc
doi: 10.1016/j.jsb.2021.107808
external_id:
  isi:
  - '000720259500002'
file:
- access_level: open_access
  checksum: 6b209e4d44775d4e02b50f78982c15fa
  content_type: application/pdf
  creator: cchlebak
  date_created: 2021-11-15T13:11:27Z
  date_updated: 2021-11-15T13:11:27Z
  file_id: '10291'
  file_name: 2021_JournalStructBiol_Dimchev.pdf
  file_size: 16818304
  relation: main_file
  success: 1
file_date_updated: 2021-11-15T13:11:27Z
has_accepted_license: '1'
intvolume: '       213'
isi: 1
issue: '4'
keyword:
- Structural Biology
language:
- iso: eng
month: '11'
oa: 1
oa_version: Published Version
project:
- _id: 9B954C5C-BA93-11EA-9121-9846C619BF3A
  grant_number: P33367
  name: Structure and isoform diversity of the Arp2/3 complex
- _id: 2674F658-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: M02495
  name: Protein structure and function in filopodia across scales
publication: Journal of Structural Biology
publication_identifier:
  issn:
  - 1047-8477
publication_status: published
publisher: 'Elsevier '
quality_controlled: '1'
related_material:
  record:
  - id: '14502'
    relation: software
    status: public
scopus_import: '1'
status: public
title: Computational toolbox for ultrastructural quantitative analysis of filament
  networks in cryo-ET data
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: 213
year: '2021'
...
---
_id: '8586'
abstract:
- lang: eng
  text: Cryo-electron microscopy (cryo-EM) of cellular specimens provides insights
    into biological processes and structures within a native context. However, a major
    challenge still lies in the efficient and reproducible preparation of adherent
    cells for subsequent cryo-EM analysis. This is due to the sensitivity of many
    cellular specimens to the varying seeding and culturing conditions required for
    EM experiments, the often limited amount of cellular material and also the fragility
    of EM grids and their substrate. Here, we present low-cost and reusable 3D printed
    grid holders, designed to improve specimen preparation when culturing challenging
    cellular samples directly on grids. The described grid holders increase cell culture
    reproducibility and throughput, and reduce the resources required for cell culturing.
    We show that grid holders can be integrated into various cryo-EM workflows, including
    micro-patterning approaches to control cell seeding on grids, and for generating
    samples for cryo-focused ion beam milling and cryo-electron tomography experiments.
    Their adaptable design allows for the generation of specialized grid holders customized
    to a large variety of applications.
acknowledged_ssus:
- _id: ScienComp
- _id: LifeSc
- _id: Bio
- _id: EM-Fac
acknowledgement: This work was supported by the Austrian Science Fund (FWF, P33367)
  to FKMS. BZ acknowledges support by the Niederösterreich Fond. This research was
  also supported by the Scientific Service Units (SSU) of IST Austria through resources
  provided by Scientific Computing (SciComp), the Life Science Facility (LSF), the
  BioImaging Facility (BIF) and the Electron Microscopy Facility (EMF). We thank Georgi
  Dimchev (IST Austria) and Sonja Jacob (Vienna Biocenter Core Facilities) for testing
  our grid holders in different experimental setups and Daniel Gütl and the Kondrashov
  group (IST Austria) for granting us repeated access to their 3D printers. We also
  thank Jonna Alanko and the Sixt lab (IST Austria) for providing us HeLa cells, primary
  BL6 mouse tail fibroblasts, NIH 3T3 fibroblasts and human telomerase immortalised
  foreskin fibroblasts for our experiments. We are thankful to Ori Avinoam and William
  Wan for helpful comments on the manuscript and also thank Dorotea Fracchiolla (Art&Science)
  for illustrating the graphical abstract.
article_number: '107633'
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Florian
  full_name: Fäßler, Florian
  id: 404F5528-F248-11E8-B48F-1D18A9856A87
  last_name: Fäßler
  orcid: 0000-0001-7149-769X
- first_name: Bettina
  full_name: Zens, Bettina
  id: 45FD126C-F248-11E8-B48F-1D18A9856A87
  last_name: Zens
- first_name: Robert
  full_name: Hauschild, Robert
  id: 4E01D6B4-F248-11E8-B48F-1D18A9856A87
  last_name: Hauschild
  orcid: 0000-0001-9843-3522
- 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: Fäßler F, Zens B, Hauschild R, Schur FK. 3D printed cell culture grid holders
    for improved cellular specimen preparation in cryo-electron microscopy. <i>Journal
    of Structural Biology</i>. 2020;212(3). doi:<a href="https://doi.org/10.1016/j.jsb.2020.107633">10.1016/j.jsb.2020.107633</a>
  apa: Fäßler, F., Zens, B., Hauschild, R., &#38; Schur, F. K. (2020). 3D printed
    cell culture grid holders for improved cellular specimen preparation in cryo-electron
    microscopy. <i>Journal of Structural Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jsb.2020.107633">https://doi.org/10.1016/j.jsb.2020.107633</a>
  chicago: Fäßler, Florian, Bettina Zens, Robert Hauschild, and Florian KM Schur.
    “3D Printed Cell Culture Grid Holders for Improved Cellular Specimen Preparation
    in Cryo-Electron Microscopy.” <i>Journal of Structural Biology</i>. Elsevier,
    2020. <a href="https://doi.org/10.1016/j.jsb.2020.107633">https://doi.org/10.1016/j.jsb.2020.107633</a>.
  ieee: F. Fäßler, B. Zens, R. Hauschild, and F. K. Schur, “3D printed cell culture
    grid holders for improved cellular specimen preparation in cryo-electron microscopy,”
    <i>Journal of Structural Biology</i>, vol. 212, no. 3. Elsevier, 2020.
  ista: Fäßler F, Zens B, Hauschild R, Schur FK. 2020. 3D printed cell culture grid
    holders for improved cellular specimen preparation in cryo-electron microscopy.
    Journal of Structural Biology. 212(3), 107633.
  mla: Fäßler, Florian, et al. “3D Printed Cell Culture Grid Holders for Improved
    Cellular Specimen Preparation in Cryo-Electron Microscopy.” <i>Journal of Structural
    Biology</i>, vol. 212, no. 3, 107633, Elsevier, 2020, doi:<a href="https://doi.org/10.1016/j.jsb.2020.107633">10.1016/j.jsb.2020.107633</a>.
  short: F. Fäßler, B. Zens, R. Hauschild, F.K. Schur, Journal of Structural Biology
    212 (2020).
date_created: 2020-09-29T13:24:06Z
date_published: 2020-12-01T00:00:00Z
date_updated: 2024-03-25T23:30:04Z
day: '01'
ddc:
- '570'
department:
- _id: FlSc
doi: 10.1016/j.jsb.2020.107633
external_id:
  isi:
  - '000600997800008'
file:
- access_level: open_access
  checksum: c48cbf594e84fc2f91966ffaafc0918c
  content_type: application/pdf
  creator: dernst
  date_created: 2020-12-10T14:01:10Z
  date_updated: 2020-12-10T14:01:10Z
  file_id: '8937'
  file_name: 2020_JourStrucBiology_Faessler.pdf
  file_size: 7076870
  relation: main_file
  success: 1
file_date_updated: 2020-12-10T14:01:10Z
has_accepted_license: '1'
intvolume: '       212'
isi: 1
issue: '3'
keyword:
- electron microscopy
- cryo-EM
- EM sample preparation
- 3D printing
- cell culture
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
project:
- _id: 9B954C5C-BA93-11EA-9121-9846C619BF3A
  grant_number: P33367
  name: Structure and isoform diversity of the Arp2/3 complex
- _id: 059B463C-7A3F-11EA-A408-12923DDC885E
  name: NÖ-Fonds Preis für die Jungforscherin des Jahres am IST Austria
publication: Journal of Structural Biology
publication_identifier:
  issn:
  - 1047-8477
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  record:
  - id: '14592'
    relation: used_in_publication
    status: public
  - id: '12491'
    relation: dissertation_contains
    status: public
scopus_import: '1'
status: public
title: 3D printed cell culture grid holders for improved cellular specimen preparation
  in cryo-electron 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: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 212
year: '2020'
...
---
_id: '8409'
abstract:
- lang: eng
  text: The bacterial cell wall is composed of the peptidoglycan (PG), a large polymer
    that maintains the integrity of the bacterial cell. Due to its multi-gigadalton
    size, heterogeneity, and dynamics, atomic-resolution studies are inherently complex.
    Solid-state NMR is an important technique to gain insight into its structure,
    dynamics and interactions. Here, we explore the possibilities to study the PG
    with ultra-fast (100 kHz) magic-angle spinning NMR. We demonstrate that highly
    resolved spectra can be obtained, and show strategies to obtain site-specific
    resonance assignments and distance information. We also explore the use of proton-proton
    correlation experiments, thus opening the way for NMR studies of intact cell walls
    without the need for isotope labeling.
article_processing_charge: No
article_type: original
author:
- first_name: Catherine
  full_name: Bougault, Catherine
  last_name: Bougault
- first_name: Isabel
  full_name: Ayala, Isabel
  last_name: Ayala
- first_name: Waldemar
  full_name: Vollmer, Waldemar
  last_name: Vollmer
- first_name: Jean-Pierre
  full_name: Simorre, Jean-Pierre
  last_name: Simorre
- first_name: Paul
  full_name: Schanda, Paul
  id: 7B541462-FAF6-11E9-A490-E8DFE5697425
  last_name: Schanda
  orcid: 0000-0002-9350-7606
citation:
  ama: Bougault C, Ayala I, Vollmer W, Simorre J-P, Schanda P. Studying intact bacterial
    peptidoglycan by proton-detected NMR spectroscopy at 100 kHz MAS frequency. <i>Journal
    of Structural Biology</i>. 2019;206(1):66-72. doi:<a href="https://doi.org/10.1016/j.jsb.2018.07.009">10.1016/j.jsb.2018.07.009</a>
  apa: Bougault, C., Ayala, I., Vollmer, W., Simorre, J.-P., &#38; Schanda, P. (2019).
    Studying intact bacterial peptidoglycan by proton-detected NMR spectroscopy at
    100 kHz MAS frequency. <i>Journal of Structural Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jsb.2018.07.009">https://doi.org/10.1016/j.jsb.2018.07.009</a>
  chicago: Bougault, Catherine, Isabel Ayala, Waldemar Vollmer, Jean-Pierre Simorre,
    and Paul Schanda. “Studying Intact Bacterial Peptidoglycan by Proton-Detected
    NMR Spectroscopy at 100 kHz MAS Frequency.” <i>Journal of Structural Biology</i>.
    Elsevier, 2019. <a href="https://doi.org/10.1016/j.jsb.2018.07.009">https://doi.org/10.1016/j.jsb.2018.07.009</a>.
  ieee: C. Bougault, I. Ayala, W. Vollmer, J.-P. Simorre, and P. Schanda, “Studying
    intact bacterial peptidoglycan by proton-detected NMR spectroscopy at 100 kHz
    MAS frequency,” <i>Journal of Structural Biology</i>, vol. 206, no. 1. Elsevier,
    pp. 66–72, 2019.
  ista: Bougault C, Ayala I, Vollmer W, Simorre J-P, Schanda P. 2019. Studying intact
    bacterial peptidoglycan by proton-detected NMR spectroscopy at 100 kHz MAS frequency.
    Journal of Structural Biology. 206(1), 66–72.
  mla: Bougault, Catherine, et al. “Studying Intact Bacterial Peptidoglycan by Proton-Detected
    NMR Spectroscopy at 100 kHz MAS Frequency.” <i>Journal of Structural Biology</i>,
    vol. 206, no. 1, Elsevier, 2019, pp. 66–72, doi:<a href="https://doi.org/10.1016/j.jsb.2018.07.009">10.1016/j.jsb.2018.07.009</a>.
  short: C. Bougault, I. Ayala, W. Vollmer, J.-P. Simorre, P. Schanda, Journal of
    Structural Biology 206 (2019) 66–72.
date_created: 2020-09-17T10:29:10Z
date_published: 2019-04-01T00:00:00Z
date_updated: 2021-01-12T08:19:05Z
day: '01'
doi: 10.1016/j.jsb.2018.07.009
extern: '1'
external_id:
  pmid:
  - '30031884'
intvolume: '       206'
issue: '1'
keyword:
- Structural Biology
language:
- iso: eng
month: '04'
oa_version: Submitted Version
page: 66-72
pmid: 1
publication: Journal of Structural Biology
publication_identifier:
  issn:
  - 1047-8477
publication_status: published
publisher: Elsevier
quality_controlled: '1'
status: public
title: Studying intact bacterial peptidoglycan by proton-detected NMR spectroscopy
  at 100 kHz MAS frequency
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 206
year: '2019'
...
---
_id: '9655'
abstract:
- lang: eng
  text: Correlative microscopy incorporates the specificity of fluorescent protein
    labeling into high-resolution electron micrographs. Several approaches exist for
    correlative microscopy, most of which have used the green fluorescent protein
    (GFP) as the label for light microscopy. Here we use chemical tagging and synthetic
    fluorophores instead, in order to achieve protein-specific labeling, and to perform
    multicolor imaging. We show that synthetic fluorophores preserve their post-embedding
    fluorescence in the presence of uranyl acetate. Post-embedding fluorescence is
    of such quality that the specimen can be prepared with identical protocols for
    scanning electron microscopy (SEM) and transmission electron microscopy (TEM);
    this is particularly valuable when singular or otherwise difficult samples are
    examined. We show that synthetic fluorophores give bright, well-resolved signals
    in super-resolution light microscopy, enabling us to superimpose light microscopic
    images with a precision of up to 25 nm in the x–y plane on electron micrographs.
    To exemplify the preservation quality of our new method we visualize the molecular
    arrangement of cadherins in adherens junctions of mouse epithelial cells.
article_processing_charge: No
article_type: original
author:
- first_name: Mario
  full_name: Perkovic, Mario
  last_name: Perkovic
- first_name: Michael
  full_name: Kunz, Michael
  last_name: Kunz
- first_name: Ulrike
  full_name: Endesfelder, Ulrike
  last_name: Endesfelder
- first_name: Stefanie
  full_name: Bunse, Stefanie
  last_name: Bunse
- first_name: Christoph
  full_name: Wigge, Christoph
  last_name: Wigge
- first_name: Zhou
  full_name: Yu, Zhou
  last_name: Yu
- first_name: Victor-Valentin
  full_name: Hodirnau, Victor-Valentin
  id: 3661B498-F248-11E8-B48F-1D18A9856A87
  last_name: Hodirnau
- first_name: Margot P.
  full_name: Scheffer, Margot P.
  last_name: Scheffer
- first_name: Anja
  full_name: Seybert, Anja
  last_name: Seybert
- first_name: Sebastian
  full_name: Malkusch, Sebastian
  last_name: Malkusch
- first_name: Erin M.
  full_name: Schuman, Erin M.
  last_name: Schuman
- first_name: Mike
  full_name: Heilemann, Mike
  last_name: Heilemann
- first_name: Achilleas S.
  full_name: Frangakis, Achilleas S.
  last_name: Frangakis
citation:
  ama: Perkovic M, Kunz M, Endesfelder U, et al. Correlative light- and electron microscopy
    with chemical tags. <i>Journal of Structural Biology</i>. 2014;186(2):205-213.
    doi:<a href="https://doi.org/10.1016/j.jsb.2014.03.018">10.1016/j.jsb.2014.03.018</a>
  apa: Perkovic, M., Kunz, M., Endesfelder, U., Bunse, S., Wigge, C., Yu, Z., … Frangakis,
    A. S. (2014). Correlative light- and electron microscopy with chemical tags. <i>Journal
    of Structural Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jsb.2014.03.018">https://doi.org/10.1016/j.jsb.2014.03.018</a>
  chicago: Perkovic, Mario, Michael Kunz, Ulrike Endesfelder, Stefanie Bunse, Christoph
    Wigge, Zhou Yu, Victor-Valentin Hodirnau, et al. “Correlative Light- and Electron
    Microscopy with Chemical Tags.” <i>Journal of Structural Biology</i>. Elsevier,
    2014. <a href="https://doi.org/10.1016/j.jsb.2014.03.018">https://doi.org/10.1016/j.jsb.2014.03.018</a>.
  ieee: M. Perkovic <i>et al.</i>, “Correlative light- and electron microscopy with
    chemical tags,” <i>Journal of Structural Biology</i>, vol. 186, no. 2. Elsevier,
    pp. 205–213, 2014.
  ista: Perkovic M, Kunz M, Endesfelder U, Bunse S, Wigge C, Yu Z, Hodirnau V-V, Scheffer
    MP, Seybert A, Malkusch S, Schuman EM, Heilemann M, Frangakis AS. 2014. Correlative
    light- and electron microscopy with chemical tags. Journal of Structural Biology.
    186(2), 205–213.
  mla: Perkovic, Mario, et al. “Correlative Light- and Electron Microscopy with Chemical
    Tags.” <i>Journal of Structural Biology</i>, vol. 186, no. 2, Elsevier, 2014,
    pp. 205–13, doi:<a href="https://doi.org/10.1016/j.jsb.2014.03.018">10.1016/j.jsb.2014.03.018</a>.
  short: M. Perkovic, M. Kunz, U. Endesfelder, S. Bunse, C. Wigge, Z. Yu, V.-V. Hodirnau,
    M.P. Scheffer, A. Seybert, S. Malkusch, E.M. Schuman, M. Heilemann, A.S. Frangakis,
    Journal of Structural Biology 186 (2014) 205–213.
date_created: 2021-07-14T09:05:42Z
date_published: 2014-05-01T00:00:00Z
date_updated: 2021-07-22T08:26:32Z
day: '01'
ddc:
- '570'
doi: 10.1016/j.jsb.2014.03.018
extern: '1'
external_id:
  pmid:
  - '24698954'
file:
- access_level: open_access
  checksum: a322991b43cdc5935c99db88d285aa3a
  content_type: application/pdf
  creator: asandaue
  date_created: 2021-07-22T08:06:34Z
  date_updated: 2021-07-22T08:06:34Z
  file_id: '9701'
  file_name: 2014_JournalOfStructuralBiology_Perkovic.pdf
  file_size: 3454628
  relation: main_file
  success: 1
file_date_updated: 2021-07-22T08:06:34Z
has_accepted_license: '1'
intvolume: '       186'
issue: '2'
language:
- iso: eng
license: https://creativecommons.org/licenses/by-nc-nd/3.0/
month: '05'
oa: 1
oa_version: Published Version
page: 205-213
pmid: 1
publication: Journal of Structural Biology
publication_identifier:
  issn:
  - 1047-8477
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Correlative light- and electron microscopy with chemical tags
tmp:
  image: /images/cc_by_nc_nd.png
  legal_code_url: https://creativecommons.org/licenses/by-nc-nd/3.0/legalcode
  name: Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported (CC BY-NC-ND
    3.0)
  short: CC BY-NC-ND (3.0)
type: journal_article
user_id: 6785fbc1-c503-11eb-8a32-93094b40e1cf
volume: 186
year: '2014'
...