---
_id: '6052'
abstract:
- lang: eng
  text: 'Expansion microscopy is a relatively new approach to super-resolution imaging
    that uses expandable hydrogels to isotropically increase the physical distance
    between fluorophores in biological samples such as cell cultures or tissue slices.
    The classic gel recipe results in an expansion factor of ~4×, with a resolution
    of 60–80 nm. We have recently developed X10 microscopy, which uses a gel that
    achieves an expansion factor of ~10×, with a resolution of ~25 nm. Here, we provide
    a step-by-step protocol for X10 expansion microscopy. A typical experiment consists
    of seven sequential stages: (i) immunostaining, (ii) anchoring, (iii) polymerization,
    (iv) homogenization, (v) expansion, (vi) imaging, and (vii) validation. The protocol
    presented here includes recommendations for optimization, pitfalls and their solutions,
    and detailed guidelines that should increase reproducibility. Although our protocol
    focuses on X10 expansion microscopy, we detail which of these suggestions are
    also applicable to classic fourfold expansion microscopy. We exemplify our protocol
    using primary hippocampal neurons from rats, but our approach can be used with
    other primary cells or cultured cell lines of interest. This protocol will enable
    any researcher with basic experience in immunostainings and access to an epifluorescence
    microscope to perform super-resolution microscopy with X10. The procedure takes
    3 d and requires ~5 h of actively handling the sample for labeling and expansion,
    and another ~3 h for imaging and analysis.'
article_processing_charge: No
article_type: original
author:
- first_name: Sven M
  full_name: Truckenbrodt, Sven M
  id: 45812BD4-F248-11E8-B48F-1D18A9856A87
  last_name: Truckenbrodt
- first_name: Christoph M
  full_name: Sommer, Christoph M
  id: 4DF26D8C-F248-11E8-B48F-1D18A9856A87
  last_name: Sommer
  orcid: 0000-0003-1216-9105
- first_name: Silvio O
  full_name: Rizzoli, Silvio O
  last_name: Rizzoli
- 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: Truckenbrodt SM, Sommer CM, Rizzoli SO, Danzl JG. A practical guide to optimization
    in X10 expansion microscopy. <i>Nature Protocols</i>. 2019;14(3):832–863. doi:<a
    href="https://doi.org/10.1038/s41596-018-0117-3">10.1038/s41596-018-0117-3</a>
  apa: Truckenbrodt, S. M., Sommer, C. M., Rizzoli, S. O., &#38; Danzl, J. G. (2019).
    A practical guide to optimization in X10 expansion microscopy. <i>Nature Protocols</i>.
    Nature Publishing Group. <a href="https://doi.org/10.1038/s41596-018-0117-3">https://doi.org/10.1038/s41596-018-0117-3</a>
  chicago: Truckenbrodt, Sven M, Christoph M Sommer, Silvio O Rizzoli, and Johann
    G Danzl. “A Practical Guide to Optimization in X10 Expansion Microscopy.” <i>Nature
    Protocols</i>. Nature Publishing Group, 2019. <a href="https://doi.org/10.1038/s41596-018-0117-3">https://doi.org/10.1038/s41596-018-0117-3</a>.
  ieee: S. M. Truckenbrodt, C. M. Sommer, S. O. Rizzoli, and J. G. Danzl, “A practical
    guide to optimization in X10 expansion microscopy,” <i>Nature Protocols</i>, vol.
    14, no. 3. Nature Publishing Group, pp. 832–863, 2019.
  ista: Truckenbrodt SM, Sommer CM, Rizzoli SO, Danzl JG. 2019. A practical guide
    to optimization in X10 expansion microscopy. Nature Protocols. 14(3), 832–863.
  mla: Truckenbrodt, Sven M., et al. “A Practical Guide to Optimization in X10 Expansion
    Microscopy.” <i>Nature Protocols</i>, vol. 14, no. 3, Nature Publishing Group,
    2019, pp. 832–863, doi:<a href="https://doi.org/10.1038/s41596-018-0117-3">10.1038/s41596-018-0117-3</a>.
  short: S.M. Truckenbrodt, C.M. Sommer, S.O. Rizzoli, J.G. Danzl, Nature Protocols
    14 (2019) 832–863.
date_created: 2019-02-24T22:59:20Z
date_published: 2019-03-01T00:00:00Z
date_updated: 2025-04-14T07:44:00Z
day: '01'
ddc:
- '570'
department:
- _id: JoDa
- _id: Bio
doi: 10.1038/s41596-018-0117-3
ec_funded: 1
external_id:
  isi:
  - '000459890700008'
  pmid:
  - '30778205'
file:
- access_level: open_access
  checksum: 7efb9951e7ddf3e3dcc2fb92b859c623
  content_type: application/vnd.openxmlformats-officedocument.wordprocessingml.document
  creator: kschuh
  date_created: 2021-06-29T14:41:46Z
  date_updated: 2021-06-29T14:41:46Z
  file_id: '9619'
  file_name: 181031_Truckenbrodt_ExM_NatProtoc.docx
  file_size: 84478958
  relation: main_file
  success: 1
file_date_updated: 2021-06-29T14:41:46Z
has_accepted_license: '1'
intvolume: '        14'
isi: 1
issue: '3'
language:
- iso: eng
month: '03'
oa: 1
oa_version: Submitted Version
page: 832–863
pmid: 1
project:
- _id: 260C2330-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '754411'
  name: ISTplus - Postdoctoral Fellowships
- _id: 265CB4D0-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: I03600
  name: Optical control of synaptic function via adhesion molecules
publication: Nature Protocols
publication_status: published
publisher: Nature Publishing Group
quality_controlled: '1'
scopus_import: '1'
status: public
title: A practical guide to optimization in X10 expansion microscopy
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 14
year: '2019'
...