---
_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: 2023-10-17T08:42:24Z
day: '01'
department:
- _id: RySh
- _id: EM-Fac
doi: 10.1369/0022155418786698
external_id:
  isi:
  - '000452277700005'
  pmid:
  - '29969056'
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'
...