---
_id: '14794'
abstract:
- lang: eng
  text: "Mosaic analysis with double markers (MADM) technology enables the sparse
    labeling of genetically defined neurons. We present a protocol for time-lapse
    imaging of cortical projection neuron migration in mice using MADM. We describe
    steps for the isolation, culturing, and 4D imaging of neuronal dynamics in MADM-labeled
    brain tissue. While this protocol is compatible with other single-cell labeling
    methods, the MADM approach provides a genetic platform for the functional assessment
    of cell-autonomous candidate gene function and the relative contribution of non-cell-autonomous
    effects.\r\n\r\nFor complete details on the use and execution of this protocol,
    please refer to Hansen et al. (2022),1 Contreras et al. (2021),2 and Amberg and
    Hippenmeyer (2021).3"
acknowledged_ssus:
- _id: Bio
- _id: PreCl
acknowledgement: We thank Florian Pauler for discussion and his expert technical support.
  This research was supported by the Scientific Service Units (SSU) at IST Austria
  through resources provided by the Imaging and Optics Facility (IOF) and Preclinical
  Facility (PCF). A.H.H. was a recipient of a DOC Fellowship (24812) of the Austrian
  Academy of Sciences.
article_number: '102795'
article_processing_charge: Yes
article_type: review
author:
- first_name: Andi H
  full_name: Hansen, Andi H
  id: 38853E16-F248-11E8-B48F-1D18A9856A87
  last_name: Hansen
- first_name: Simon
  full_name: Hippenmeyer, Simon
  id: 37B36620-F248-11E8-B48F-1D18A9856A87
  last_name: Hippenmeyer
  orcid: 0000-0003-2279-1061
citation:
  ama: Hansen AH, Hippenmeyer S. Time-lapse imaging of cortical projection neuron
    migration in mice using mosaic analysis with double markers. <i>STAR Protocols</i>.
    2024;5(1). doi:<a href="https://doi.org/10.1016/j.xpro.2023.102795">10.1016/j.xpro.2023.102795</a>
  apa: Hansen, A. H., &#38; Hippenmeyer, S. (2024). Time-lapse imaging of cortical
    projection neuron migration in mice using mosaic analysis with double markers.
    <i>STAR Protocols</i>. Elsevier. <a href="https://doi.org/10.1016/j.xpro.2023.102795">https://doi.org/10.1016/j.xpro.2023.102795</a>
  chicago: Hansen, Andi H, and Simon Hippenmeyer. “Time-Lapse Imaging of Cortical
    Projection Neuron Migration in Mice Using Mosaic Analysis with Double Markers.”
    <i>STAR Protocols</i>. Elsevier, 2024. <a href="https://doi.org/10.1016/j.xpro.2023.102795">https://doi.org/10.1016/j.xpro.2023.102795</a>.
  ieee: A. H. Hansen and S. Hippenmeyer, “Time-lapse imaging of cortical projection
    neuron migration in mice using mosaic analysis with double markers,” <i>STAR Protocols</i>,
    vol. 5, no. 1. Elsevier, 2024.
  ista: Hansen AH, Hippenmeyer S. 2024. Time-lapse imaging of cortical projection
    neuron migration in mice using mosaic analysis with double markers. STAR Protocols.
    5(1), 102795.
  mla: Hansen, Andi H., and Simon Hippenmeyer. “Time-Lapse Imaging of Cortical Projection
    Neuron Migration in Mice Using Mosaic Analysis with Double Markers.” <i>STAR Protocols</i>,
    vol. 5, no. 1, 102795, Elsevier, 2024, doi:<a href="https://doi.org/10.1016/j.xpro.2023.102795">10.1016/j.xpro.2023.102795</a>.
  short: A.H. Hansen, S. Hippenmeyer, STAR Protocols 5 (2024).
date_created: 2024-01-14T23:00:56Z
date_published: 2024-01-01T00:00:00Z
date_updated: 2025-08-11T11:49:30Z
day: '01'
department:
- _id: SiHi
doi: 10.1016/j.xpro.2023.102795
external_id:
  oaworkID:
  - '34426698 '
  pmid:
  - '38165800'
intvolume: '         5'
issue: '1'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: https://doi.org/10.1016/j.xpro.2023.102795
month: '01'
oa: 1
oa_version: Published Version
oaworkID: 1
pmid: 1
project:
- _id: 2625A13E-B435-11E9-9278-68D0E5697425
  grant_number: '24812'
  name: Molecular Mechanisms of Radial Neuronal Migration
publication: STAR Protocols
publication_identifier:
  eissn:
  - 2666-1667
publication_status: epub_ahead
publisher: Elsevier
quality_controlled: '1'
related_material:
  link:
  - relation: software
    url: http://github.com/hippenmeyerlab
scopus_import: '1'
status: public
title: Time-lapse imaging of cortical projection neuron migration in mice using mosaic
  analysis with double markers
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 5
year: '2024'
...
---
_id: '10321'
abstract:
- lang: eng
  text: Mosaic analysis with double markers (MADM) technology enables the generation
    of genetic mosaic tissue in mice. MADM enables concomitant fluorescent cell labeling
    and introduction of a mutation of a gene of interest with single-cell resolution.
    This protocol highlights major steps for the generation of genetic mosaic tissue
    and the isolation and processing of respective tissues for downstream histological
    analysis. For complete details on the use and execution of this protocol, please
    refer to Contreras et al. (2021).
acknowledged_ssus:
- _id: Bio
- _id: PreCl
acknowledgement: This research was supported by the Scientific Service Units (SSU)
  at IST Austria through resources provided by the Bioimaging (BIF) and Preclinical
  Facilities (PCF). We particularly thank Mohammad Goudarzi for assistance with photography
  of mouse perfusion and dissection. N.A. received support from FWF Firnberg-Programm
  (T 1031). This work was also supported by IST Austria institutional funds; FWF SFB
  F78 to S.H.; and the European Research Council (ERC) under the European Union’s
  Horizon 2020 research and innovation programme (grant agreement no. 725780 LinPro)
  to S.H.
article_number: '100939'
article_processing_charge: Yes
article_type: original
author:
- first_name: Nicole
  full_name: Amberg, Nicole
  id: 4CD6AAC6-F248-11E8-B48F-1D18A9856A87
  last_name: Amberg
  orcid: 0000-0002-3183-8207
- first_name: Simon
  full_name: Hippenmeyer, Simon
  id: 37B36620-F248-11E8-B48F-1D18A9856A87
  last_name: Hippenmeyer
  orcid: 0000-0003-2279-1061
citation:
  ama: Amberg N, Hippenmeyer S. Genetic mosaic dissection of candidate genes in mice
    using mosaic analysis with double markers. <i>STAR Protocols</i>. 2021;2(4). doi:<a
    href="https://doi.org/10.1016/j.xpro.2021.100939">10.1016/j.xpro.2021.100939</a>
  apa: Amberg, N., &#38; Hippenmeyer, S. (2021). Genetic mosaic dissection of candidate
    genes in mice using mosaic analysis with double markers. <i>STAR Protocols</i>.
    Cell Press. <a href="https://doi.org/10.1016/j.xpro.2021.100939">https://doi.org/10.1016/j.xpro.2021.100939</a>
  chicago: Amberg, Nicole, and Simon Hippenmeyer. “Genetic Mosaic Dissection of Candidate
    Genes in Mice Using Mosaic Analysis with Double Markers.” <i>STAR Protocols</i>.
    Cell Press, 2021. <a href="https://doi.org/10.1016/j.xpro.2021.100939">https://doi.org/10.1016/j.xpro.2021.100939</a>.
  ieee: N. Amberg and S. Hippenmeyer, “Genetic mosaic dissection of candidate genes
    in mice using mosaic analysis with double markers,” <i>STAR Protocols</i>, vol.
    2, no. 4. Cell Press, 2021.
  ista: Amberg N, Hippenmeyer S. 2021. Genetic mosaic dissection of candidate genes
    in mice using mosaic analysis with double markers. STAR Protocols. 2(4), 100939.
  mla: Amberg, Nicole, and Simon Hippenmeyer. “Genetic Mosaic Dissection of Candidate
    Genes in Mice Using Mosaic Analysis with Double Markers.” <i>STAR Protocols</i>,
    vol. 2, no. 4, 100939, Cell Press, 2021, doi:<a href="https://doi.org/10.1016/j.xpro.2021.100939">10.1016/j.xpro.2021.100939</a>.
  short: N. Amberg, S. Hippenmeyer, STAR Protocols 2 (2021).
date_created: 2021-11-21T23:01:28Z
date_published: 2021-11-10T00:00:00Z
date_updated: 2023-11-16T13:08:03Z
day: '10'
ddc:
- '573'
department:
- _id: SiHi
doi: 10.1016/j.xpro.2021.100939
ec_funded: 1
file:
- access_level: open_access
  checksum: 9e3f6d06bf583e7a8b6a9e9a60500a28
  content_type: application/pdf
  creator: cchlebak
  date_created: 2021-11-22T08:23:58Z
  date_updated: 2021-11-22T08:23:58Z
  file_id: '10329'
  file_name: 2021_STARProtocols_Amberg.pdf
  file_size: 7309464
  relation: main_file
  success: 1
file_date_updated: 2021-11-22T08:23:58Z
has_accepted_license: '1'
intvolume: '         2'
issue: '4'
language:
- iso: eng
month: '11'
oa: 1
oa_version: Published Version
project:
- _id: 260018B0-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '725780'
  name: Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development
- _id: 268F8446-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: T0101031
  name: Role of Eed in neural stem cell lineage progression
- _id: 059F6AB4-7A3F-11EA-A408-12923DDC885E
  grant_number: F07805
  name: Molecular Mechanisms of Neural Stem Cell Lineage Progression
publication: STAR Protocols
publication_identifier:
  eissn:
  - 2666-1667
publication_status: published
publisher: Cell Press
quality_controlled: '1'
scopus_import: '1'
status: public
title: Genetic mosaic dissection of candidate genes in mice using mosaic analysis
  with double markers
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: 3E5EF7F0-F248-11E8-B48F-1D18A9856A87
volume: 2
year: '2021'
...
---
_id: '10565'
abstract:
- lang: eng
  text: 'Enzymatic digestion of the extracellular matrix with chondroitinase-ABC reinstates
    juvenile-like plasticity in the adult cortex as it also disassembles the perineuronal
    nets (PNNs). The disadvantage of the enzyme is that it must be applied intracerebrally
    and it degrades the ECM for several weeks. Here, we provide two minimally invasive
    and transient protocols for microglia-enabled PNN disassembly in mouse cortex:
    repeated treatment with ketamine-xylazine-acepromazine (KXA) anesthesia and 60-Hz
    light entrainment. We also discuss how to analyze PNNs within microglial endosomes-lysosomes.
    For complete details on the use and execution of this protocol, please refer to
    Venturino et al. (2021).'
acknowledged_ssus:
- _id: Bio
acknowledgement: This research was supported by the European Research Council (grant
  715571 to S.S.). We thank Rouven Schulz, Michael Schunn, Claudia Gold, Gabriel Krens,
  Sarah Gorkiewicz, Margaret Maes, Jürgen Siegert, Marco Benevento, and Sara Oakeley
  for comments on the manuscript and the IST Austria Bioimaging Facility for the technical
  support.
article_number: '101012'
article_processing_charge: Yes
article_type: original
author:
- first_name: Alessandro
  full_name: Venturino, Alessandro
  id: 41CB84B2-F248-11E8-B48F-1D18A9856A87
  last_name: Venturino
  orcid: 0000-0003-2356-9403
- first_name: Sandra
  full_name: Siegert, Sandra
  id: 36ACD32E-F248-11E8-B48F-1D18A9856A87
  last_name: Siegert
  orcid: 0000-0001-8635-0877
citation:
  ama: Venturino A, Siegert S. Minimally invasive protocols and quantification for
    microglia-mediated perineuronal net disassembly in mouse brain. <i>STAR Protocols</i>.
    2021;2(4). doi:<a href="https://doi.org/10.1016/j.xpro.2021.101012">10.1016/j.xpro.2021.101012</a>
  apa: Venturino, A., &#38; Siegert, S. (2021). Minimally invasive protocols and quantification
    for microglia-mediated perineuronal net disassembly in mouse brain. <i>STAR Protocols</i>.
    Elsevier ; Cell Press. <a href="https://doi.org/10.1016/j.xpro.2021.101012">https://doi.org/10.1016/j.xpro.2021.101012</a>
  chicago: Venturino, Alessandro, and Sandra Siegert. “Minimally Invasive Protocols
    and Quantification for Microglia-Mediated Perineuronal Net Disassembly in Mouse
    Brain.” <i>STAR Protocols</i>. Elsevier ; Cell Press, 2021. <a href="https://doi.org/10.1016/j.xpro.2021.101012">https://doi.org/10.1016/j.xpro.2021.101012</a>.
  ieee: A. Venturino and S. Siegert, “Minimally invasive protocols and quantification
    for microglia-mediated perineuronal net disassembly in mouse brain,” <i>STAR Protocols</i>,
    vol. 2, no. 4. Elsevier ; Cell Press, 2021.
  ista: Venturino A, Siegert S. 2021. Minimally invasive protocols and quantification
    for microglia-mediated perineuronal net disassembly in mouse brain. STAR Protocols.
    2(4), 101012.
  mla: Venturino, Alessandro, and Sandra Siegert. “Minimally Invasive Protocols and
    Quantification for Microglia-Mediated Perineuronal Net Disassembly in Mouse Brain.”
    <i>STAR Protocols</i>, vol. 2, no. 4, 101012, Elsevier ; Cell Press, 2021, doi:<a
    href="https://doi.org/10.1016/j.xpro.2021.101012">10.1016/j.xpro.2021.101012</a>.
  short: A. Venturino, S. Siegert, STAR Protocols 2 (2021).
date_created: 2021-12-19T23:01:32Z
date_published: 2021-12-17T00:00:00Z
date_updated: 2023-11-16T13:11:04Z
day: '17'
ddc:
- '573'
department:
- _id: SaSi
doi: 10.1016/j.xpro.2021.101012
ec_funded: 1
file:
- access_level: open_access
  checksum: 9ea2501056c5df99e84726b845e9b976
  content_type: application/pdf
  creator: cchlebak
  date_created: 2021-12-20T08:58:40Z
  date_updated: 2021-12-20T08:58:40Z
  file_id: '10570'
  file_name: 2021_STARProt_Venturino.pdf
  file_size: 6207060
  relation: main_file
  success: 1
file_date_updated: 2021-12-20T08:58:40Z
has_accepted_license: '1'
intvolume: '         2'
issue: '4'
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
project:
- _id: 25D4A630-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '715571'
  name: Microglia action towards neuronal circuit formation and function in health
    and disease
publication: STAR Protocols
publication_identifier:
  eissn:
  - 2666-1667
publication_status: published
publisher: Elsevier ; Cell Press
quality_controlled: '1'
scopus_import: '1'
status: public
title: Minimally invasive protocols and quantification for microglia-mediated perineuronal
  net disassembly in mouse brain
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: 3E5EF7F0-F248-11E8-B48F-1D18A9856A87
volume: 2
year: '2021'
...