Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions

Integrin-dependent cell-extracellular matrix adhesion is essential for wound healing, embryonic development, immunity, and tissue organization. Here, we present a protocol for the imaging and quantitative analysis of integrin-dependent cell-matrix adhesions. We describe steps for cell culture; virus...

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Autores principales: van Stalborch, Anne-Marieke D., Clark, Andrew G., Sonnenberg, Arnoud, Margadant, Coert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10469561/
https://www.ncbi.nlm.nih.gov/pubmed/37616164
http://dx.doi.org/10.1016/j.xpro.2023.102473
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author van Stalborch, Anne-Marieke D.
Clark, Andrew G.
Sonnenberg, Arnoud
Margadant, Coert
author_facet van Stalborch, Anne-Marieke D.
Clark, Andrew G.
Sonnenberg, Arnoud
Margadant, Coert
author_sort van Stalborch, Anne-Marieke D.
collection PubMed
description Integrin-dependent cell-extracellular matrix adhesion is essential for wound healing, embryonic development, immunity, and tissue organization. Here, we present a protocol for the imaging and quantitative analysis of integrin-dependent cell-matrix adhesions. We describe steps for cell culture; virus preparation; lentiviral transduction; imaging with widefield, confocal, and total internal reflection fluorescence microscopy; and using a script for their quantitative analysis. We then detail procedures for analyzing adhesion dynamics by live-cell imaging and fluorescence recovery after photobleaching (FRAP). For complete details on the use and execution of this protocol, please refer to Margadant et al. (2012),(1) van der Bijl et al. (2020),(2) Amado-Azevedo et al. (2021).(3)
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spelling pubmed-104695612023-09-01 Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions van Stalborch, Anne-Marieke D. Clark, Andrew G. Sonnenberg, Arnoud Margadant, Coert STAR Protoc Protocol Integrin-dependent cell-extracellular matrix adhesion is essential for wound healing, embryonic development, immunity, and tissue organization. Here, we present a protocol for the imaging and quantitative analysis of integrin-dependent cell-matrix adhesions. We describe steps for cell culture; virus preparation; lentiviral transduction; imaging with widefield, confocal, and total internal reflection fluorescence microscopy; and using a script for their quantitative analysis. We then detail procedures for analyzing adhesion dynamics by live-cell imaging and fluorescence recovery after photobleaching (FRAP). For complete details on the use and execution of this protocol, please refer to Margadant et al. (2012),(1) van der Bijl et al. (2020),(2) Amado-Azevedo et al. (2021).(3) Elsevier 2023-08-23 /pmc/articles/PMC10469561/ /pubmed/37616164 http://dx.doi.org/10.1016/j.xpro.2023.102473 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
van Stalborch, Anne-Marieke D.
Clark, Andrew G.
Sonnenberg, Arnoud
Margadant, Coert
Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
title Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
title_full Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
title_fullStr Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
title_full_unstemmed Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
title_short Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
title_sort imaging and quantitative analysis of integrin-dependent cell-matrix adhesions
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10469561/
https://www.ncbi.nlm.nih.gov/pubmed/37616164
http://dx.doi.org/10.1016/j.xpro.2023.102473
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AT margadantcoert imagingandquantitativeanalysisofintegrindependentcellmatrixadhesions

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