---
_id: '7665'
abstract:
- lang: eng
  text: Acute brain slice preparation is a powerful experimental model for investigating
    the characteristics of synaptic function in the brain. Although brain tissue is
    usually cut at ice-cold temperature (CT) to facilitate slicing and avoid neuronal
    damage, exposure to CT causes molecular and architectural changes of synapses.
    To address these issues, we investigated ultrastructural and electrophysiological
    features of synapses in mouse acute cerebellar slices prepared at ice-cold and
    physiological temperature (PT). In the slices prepared at CT, we found significant
    spine loss and reconstruction, synaptic vesicle rearrangement and decrease in
    synaptic proteins, all of which were not detected in slices prepared at PT. Consistent
    with these structural findings, slices prepared at PT showed higher release probability.
    Furthermore, preparation at PT allows electrophysiological recording immediately
    after slicing resulting in higher detectability of long-term depression (LTD)
    after motor learning compared with that at CT. These results indicate substantial
    advantages of the slice preparation at PT for investigating synaptic functions
    in different physiological conditions.
article_number: '63'
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Kohgaku
  full_name: Eguchi, Kohgaku
  id: 2B7846DC-F248-11E8-B48F-1D18A9856A87
  last_name: Eguchi
  orcid: 0000-0002-6170-2546
- first_name: Philipp
  full_name: Velicky, Philipp
  id: 39BDC62C-F248-11E8-B48F-1D18A9856A87
  last_name: Velicky
  orcid: 0000-0002-2340-7431
- first_name: Elena
  full_name: Hollergschwandtner, Elena
  id: 3C054040-F248-11E8-B48F-1D18A9856A87
  last_name: Hollergschwandtner
- first_name: Makoto
  full_name: Itakura, Makoto
  last_name: Itakura
- first_name: Yugo
  full_name: Fukazawa, Yugo
  last_name: Fukazawa
- first_name: Johann G
  full_name: Danzl, Johann G
  id: 42EFD3B6-F248-11E8-B48F-1D18A9856A87
  last_name: Danzl
  orcid: 0000-0001-8559-3973
- first_name: Ryuichi
  full_name: Shigemoto, Ryuichi
  id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
  last_name: Shigemoto
  orcid: 0000-0001-8761-9444
citation:
  ama: Eguchi K, Velicky P, Saeckl E, et al. Advantages of acute brain slices prepared
    at physiological temperature in the characterization of synaptic functions. <i>Frontiers
    in Cellular Neuroscience</i>. 2020;14. doi:<a href="https://doi.org/10.3389/fncel.2020.00063">10.3389/fncel.2020.00063</a>
  apa: Eguchi, K., Velicky, P., Saeckl, E., Itakura, M., Fukazawa, Y., Danzl, J. G.,
    &#38; Shigemoto, R. (2020). Advantages of acute brain slices prepared at physiological
    temperature in the characterization of synaptic functions. <i>Frontiers in Cellular
    Neuroscience</i>. Frontiers Media. <a href="https://doi.org/10.3389/fncel.2020.00063">https://doi.org/10.3389/fncel.2020.00063</a>
  chicago: Eguchi, Kohgaku, Philipp Velicky, Elena Saeckl, Makoto Itakura, Yugo Fukazawa,
    Johann G Danzl, and Ryuichi Shigemoto. “Advantages of Acute Brain Slices Prepared
    at Physiological Temperature in the Characterization of Synaptic Functions.” <i>Frontiers
    in Cellular Neuroscience</i>. Frontiers Media, 2020. <a href="https://doi.org/10.3389/fncel.2020.00063">https://doi.org/10.3389/fncel.2020.00063</a>.
  ieee: K. Eguchi <i>et al.</i>, “Advantages of acute brain slices prepared at physiological
    temperature in the characterization of synaptic functions,” <i>Frontiers in Cellular
    Neuroscience</i>, vol. 14. Frontiers Media, 2020.
  ista: Eguchi K, Velicky P, Saeckl E, Itakura M, Fukazawa Y, Danzl JG, Shigemoto
    R. 2020. Advantages of acute brain slices prepared at physiological temperature
    in the characterization of synaptic functions. Frontiers in Cellular Neuroscience.
    14, 63.
  mla: Eguchi, Kohgaku, et al. “Advantages of Acute Brain Slices Prepared at Physiological
    Temperature in the Characterization of Synaptic Functions.” <i>Frontiers in Cellular
    Neuroscience</i>, vol. 14, 63, Frontiers Media, 2020, doi:<a href="https://doi.org/10.3389/fncel.2020.00063">10.3389/fncel.2020.00063</a>.
  short: K. Eguchi, P. Velicky, E. Saeckl, M. Itakura, Y. Fukazawa, J.G. Danzl, R.
    Shigemoto, Frontiers in Cellular Neuroscience 14 (2020).
corr_author: '1'
date_created: 2020-04-19T22:00:55Z
date_published: 2020-03-19T00:00:00Z
date_updated: 2025-06-12T07:16:39Z
day: '19'
ddc:
- '570'
department:
- _id: JoDa
- _id: RySh
doi: 10.3389/fncel.2020.00063
ec_funded: 1
external_id:
  isi:
  - '000525582200001'
  pmid:
  - '32265664'
file:
- access_level: open_access
  checksum: 1c145123c6f8dc3e2e4bd5a66a1ad60e
  content_type: application/pdf
  creator: dernst
  date_created: 2020-04-20T10:59:49Z
  date_updated: 2020-07-14T12:48:01Z
  file_id: '7668'
  file_name: 2020_FrontiersCellularNeurosc_Eguchi.pdf
  file_size: 9227283
  relation: main_file
file_date_updated: 2020-07-14T12:48:01Z
fulldoi: https://doi.org/10.3389/fncel.2020.00063
has_accepted_license: '1'
intvolume: '        14'
isi: 1
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '03'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 2659CC84-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '793482'
  name: 'Ultrastructural analysis of phosphoinositides in nerve terminals: distribution,
    dynamics and physiological roles in synaptic transmission'
- _id: 25CA28EA-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '694539'
  name: 'In situ analysis of single channel subunit composition in neurons: physiological
    implication in synaptic plasticity and behaviour'
- _id: 265CB4D0-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: I03600
  name: Optical control of synaptic function via adhesion molecules
- _id: B67AFEDC-15C9-11EA-A837-991A96BB2854
  name: IST Austria Open Access Fund
publication: Frontiers in Cellular Neuroscience
publication_identifier:
  issn:
  - 1662-5102
publication_status: published
publisher: Frontiers Media
quality_controlled: '1'
scopus_import: '1'
status: public
title: Advantages of acute brain slices prepared at physiological temperature in the
  characterization of synaptic functions
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: 14
year: '2020'
...
---
_id: '163'
abstract:
- lang: eng
  text: For ultrafast fixation of biological samples to avoid artifacts, high-pressure
    freezing (HPF) followed by freeze substitution (FS) is preferred over chemical
    fixation at room temperature. After HPF, samples are maintained at low temperature
    during dehydration and fixation, while avoiding damaging recrystallization. This
    is a notoriously slow process. McDonald and Webb demonstrated, in 2011, that sample
    agitation during FS dramatically reduces the necessary time. Then, in 2015, we
    (H.G. and S.R.) introduced an agitation module into the cryochamber of an automated
    FS unit and demonstrated that the preparation of algae could be shortened from
    days to a couple of hours. We argued that variability in the processing, reproducibility,
    and safety issues are better addressed using automated FS units. For dissemination,
    we started low-cost manufacturing of agitation modules for two of the most widely
    used FS units, the Automatic Freeze Substitution Systems, AFS(1) and AFS2, from
    Leica Microsystems, using three dimensional (3D)-printing of the major components.
    To test them, several labs independently used the modules on a wide variety of
    specimens that had previously been processed by manual agitation, or without agitation.
    We demonstrate that automated processing with sample agitation saves time, increases
    flexibility with respect to sample requirements and protocols, and produces data
    of at least as good quality as other approaches.
article_processing_charge: No
article_type: original
author:
- first_name: Siegfried
  full_name: Reipert, Siegfried
  last_name: Reipert
- first_name: Helmuth
  full_name: Goldammer, Helmuth
  last_name: Goldammer
- first_name: Christine
  full_name: Richardson, Christine
  last_name: Richardson
- first_name: Martin
  full_name: Goldberg, Martin
  last_name: Goldberg
- first_name: Timothy
  full_name: Hawkins, Timothy
  last_name: Hawkins
- first_name: Elena
  full_name: Hollergschwandtner, Elena
  id: 3C054040-F248-11E8-B48F-1D18A9856A87
  last_name: Hollergschwandtner
- first_name: Walter
  full_name: Kaufmann, Walter
  id: 3F99E422-F248-11E8-B48F-1D18A9856A87
  last_name: Kaufmann
  orcid: 0000-0001-9735-5315
- first_name: Sebastian
  full_name: Antreich, Sebastian
  last_name: Antreich
- first_name: York
  full_name: Stierhof, York
  last_name: Stierhof
citation:
  ama: 'Reipert S, Goldammer H, Richardson C, et al. Agitation modules: Flexible means
    to accelerate automated freeze substitution. <i>Journal of Histochemistry and
    Cytochemistry</i>. 2018;66(12):903-921. doi:<a href="https://doi.org/10.1369/0022155418786698">10.1369/0022155418786698</a>'
  apa: 'Reipert, S., Goldammer, H., Richardson, C., Goldberg, M., Hawkins, T., Saeckl,
    E., … Stierhof, Y. (2018). Agitation modules: Flexible means to accelerate automated
    freeze substitution. <i>Journal of Histochemistry and Cytochemistry</i>. SAGE
    Publications. <a href="https://doi.org/10.1369/0022155418786698">https://doi.org/10.1369/0022155418786698</a>'
  chicago: 'Reipert, Siegfried, Helmuth Goldammer, Christine Richardson, Martin Goldberg,
    Timothy Hawkins, Elena Saeckl, Walter Kaufmann, Sebastian Antreich, and York Stierhof.
    “Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution.”
    <i>Journal of Histochemistry and Cytochemistry</i>. SAGE Publications, 2018. <a
    href="https://doi.org/10.1369/0022155418786698">https://doi.org/10.1369/0022155418786698</a>.'
  ieee: 'S. Reipert <i>et al.</i>, “Agitation modules: Flexible means to accelerate
    automated freeze substitution,” <i>Journal of Histochemistry and Cytochemistry</i>,
    vol. 66, no. 12. SAGE Publications, pp. 903–921, 2018.'
  ista: 'Reipert S, Goldammer H, Richardson C, Goldberg M, Hawkins T, Saeckl E, Kaufmann
    W, Antreich S, Stierhof Y. 2018. Agitation modules: Flexible means to accelerate
    automated freeze substitution. Journal of Histochemistry and Cytochemistry. 66(12),
    903–921.'
  mla: 'Reipert, Siegfried, et al. “Agitation Modules: Flexible Means to Accelerate
    Automated Freeze Substitution.” <i>Journal of Histochemistry and Cytochemistry</i>,
    vol. 66, no. 12, SAGE Publications, 2018, pp. 903–21, doi:<a href="https://doi.org/10.1369/0022155418786698">10.1369/0022155418786698</a>.'
  short: S. Reipert, H. Goldammer, C. Richardson, M. Goldberg, T. Hawkins, E. Saeckl,
    W. Kaufmann, S. Antreich, Y. Stierhof, Journal of Histochemistry and Cytochemistry
    66 (2018) 903–921.
date_created: 2018-12-11T11:44:57Z
date_published: 2018-12-01T00:00:00Z
date_updated: 2026-06-18T17:50:00Z
day: '01'
ddc:
- '570'
department:
- _id: RySh
- _id: EM-Fac
doi: 10.1369/0022155418786698
external_id:
  isi:
  - '000452277700005'
  pmid:
  - '29969056'
fulldoi: https://doi.org/10.1369/0022155418786698
intvolume: '        66'
isi: 1
issue: '12'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: https://doi.org/10.1369/0022155418786698
month: '12'
oa: 1
oa_version: Published Version
page: 903-921
pmid: 1
publication: Journal of Histochemistry and Cytochemistry
publication_identifier:
  issn:
  - 0022-1554
publication_status: published
publisher: SAGE Publications
quality_controlled: '1'
scopus_import: '1'
status: public
title: 'Agitation modules: Flexible means to accelerate automated freeze substitution'
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 66
year: '2018'
...