---
_id: '11182'
abstract:
- lang: eng
text: Immune cells are constantly on the move through multicellular organisms to
explore and respond to pathogens and other harmful insults. While moving, immune
cells efficiently traverse microenvironments composed of tissue cells and extracellular
fibers, which together form complex environments of various porosity, stiffness,
topography, and chemical composition. In this protocol we describe experimental
procedures to investigate immune cell migration through microenvironments of heterogeneous
porosity. In particular, we describe micro-channels, micro-pillars, and collagen
networks as cell migration paths with alternative pore size choices. Employing
micro-channels or micro-pillars that divide at junctions into alternative paths
with initially differentially sized pores allows us to precisely (1) measure the
cellular translocation time through these porous path junctions, (2) quantify
the cellular preference for individual pore sizes, and (3) image cellular components
like the nucleus and the cytoskeleton. This reductionistic experimental setup
thus can elucidate how immune cells perform decisions in complex microenvironments
of various porosity like the interstitium. The setup further allows investigation
of the underlying forces of cellular squeezing and the consequences of cellular
deformation on the integrity of the cell and its organelles. As a complementary
approach that does not require any micro-engineering expertise, we describe the
usage of three-dimensional collagen networks with different pore sizes. Whereas
we here focus on dendritic cells as a model for motile immune cells, the described
protocols are versatile as they are also applicable for other immune cell types
like neutrophils and non-immune cell types such as mesenchymal and cancer cells.
In summary, we here describe protocols to identify the mechanisms and principles
of cellular probing, decision making, and squeezing during cellular movement through
microenvironments of heterogeneous porosity.
acknowledgement: "We thank Kasia Stefanowski for excellent technical assistance, and
the Core Facility Bioimaging of the Biomedical Center (BMC) of the Ludwig-Maximilian
University for excellent support. We gratefully acknowledge financial support from
the Peter Hans Hofschneider Professorship of the Stiftung Experimentelle Biomedizin
(to J.R), from the DFG (Collaborative Research Center SFB914, project A12; and Priority
Programme SPP2332, project 492014049; both to J.R) and from the LMU Institutional
Strategy LMU-Excellent within the framework of the German Excellence Initiative
(to J.R).\r\nOpen access funding enabled and organized by Projekt DEAL."
article_number: e407
article_processing_charge: No
article_type: original
author:
- first_name: Janina
full_name: Kroll, Janina
last_name: Kroll
- first_name: Mauricio J.A.
full_name: Ruiz-Fernandez, Mauricio J.A.
last_name: Ruiz-Fernandez
- first_name: Malte B.
full_name: Braun, Malte B.
last_name: Braun
- first_name: Jack
full_name: Merrin, Jack
id: 4515C308-F248-11E8-B48F-1D18A9856A87
last_name: Merrin
orcid: 0000-0001-5145-4609
- first_name: Jörg
full_name: Renkawitz, Jörg
id: 3F0587C8-F248-11E8-B48F-1D18A9856A87
last_name: Renkawitz
orcid: 0000-0003-2856-3369
citation:
ama: Kroll J, Ruiz-Fernandez MJA, Braun MB, Merrin J, Renkawitz J. Quantifying the
probing and selection of microenvironmental pores by motile immune cells. Current
Protocols. 2022;2(4). doi:10.1002/cpz1.407
apa: Kroll, J., Ruiz-Fernandez, M. J. A., Braun, M. B., Merrin, J., & Renkawitz,
J. (2022). Quantifying the probing and selection of microenvironmental pores by
motile immune cells. Current Protocols. Wiley. https://doi.org/10.1002/cpz1.407
chicago: Kroll, Janina, Mauricio J.A. Ruiz-Fernandez, Malte B. Braun, Jack Merrin,
and Jörg Renkawitz. “Quantifying the Probing and Selection of Microenvironmental
Pores by Motile Immune Cells.” Current Protocols. Wiley, 2022. https://doi.org/10.1002/cpz1.407.
ieee: J. Kroll, M. J. A. Ruiz-Fernandez, M. B. Braun, J. Merrin, and J. Renkawitz,
“Quantifying the probing and selection of microenvironmental pores by motile immune
cells,” Current Protocols, vol. 2, no. 4. Wiley, 2022.
ista: Kroll J, Ruiz-Fernandez MJA, Braun MB, Merrin J, Renkawitz J. 2022. Quantifying
the probing and selection of microenvironmental pores by motile immune cells.
Current Protocols. 2(4), e407.
mla: Kroll, Janina, et al. “Quantifying the Probing and Selection of Microenvironmental
Pores by Motile Immune Cells.” Current Protocols, vol. 2, no. 4, e407,
Wiley, 2022, doi:10.1002/cpz1.407.
short: J. Kroll, M.J.A. Ruiz-Fernandez, M.B. Braun, J. Merrin, J. Renkawitz, Current
Protocols 2 (2022).
date_created: 2022-04-17T22:01:46Z
date_published: 2022-04-05T00:00:00Z
date_updated: 2022-05-02T08:18:00Z
day: '05'
ddc:
- '570'
department:
- _id: NanoFab
doi: 10.1002/cpz1.407
external_id:
pmid:
- '35384410'
file:
- access_level: open_access
checksum: 72152d005c367777f6cf2f6a477f0d52
content_type: application/pdf
creator: dernst
date_created: 2022-05-02T08:16:10Z
date_updated: 2022-05-02T08:16:10Z
file_id: '11347'
file_name: 2022_CurrentProtocols_Kroll.pdf
file_size: 2142703
relation: main_file
success: 1
file_date_updated: 2022-05-02T08:16:10Z
has_accepted_license: '1'
intvolume: ' 2'
issue: '4'
language:
- iso: eng
month: '04'
oa: 1
oa_version: Published Version
pmid: 1
publication: Current Protocols
publication_identifier:
eissn:
- 2691-1299
publication_status: published
publisher: Wiley
quality_controlled: '1'
scopus_import: '1'
status: public
title: Quantifying the probing and selection of microenvironmental pores by motile
immune cells
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: 2
year: '2022'
...