Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique

Summary: Dermal fibroblasts play a central role in the production and remodeling of dermal extracellular matrix. Their spatially distinct subpopulations exhibit different functions during skin homeostasis, wound healing, and other diseases such as fibrosis and cancer. Here, we present a protocol for...

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Main Authors: Toru Hiratsuka, Emanuel Rognoni
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725002072
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author Toru Hiratsuka
Emanuel Rognoni
author_facet Toru Hiratsuka
Emanuel Rognoni
author_sort Toru Hiratsuka
collection DOAJ
description Summary: Dermal fibroblasts play a central role in the production and remodeling of dermal extracellular matrix. Their spatially distinct subpopulations exhibit different functions during skin homeostasis, wound healing, and other diseases such as fibrosis and cancer. Here, we present a protocol for detecting 3D dermal fibroblast cellular dynamics in mice without fixing the tissue using an intravital imaging technique. We describe steps for preparing, labeling, and mounting mice. We then describe detailed procedures for intravital imaging, recovery, and quantification.For complete details on the use and execution of this protocol, please refer to Rognoni et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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spelling doaj-art-e399f711b6ee45a4af8e7e780de0a4532025-08-20T02:16:18ZengElsevierSTAR Protocols2666-16672025-06-016210380110.1016/j.xpro.2025.103801Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging techniqueToru Hiratsuka0Emanuel Rognoni1Department of Oncogenesis and Growth Regulation, Research Center, Osaka International Cancer Institute, Chuoku, Osaka 541-8567, Japan; Department of Molcular Oncology, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan; Corresponding authorCentre for Cell Biology and Cutaneous Research, The Faculty of Medicine and Dentistry, Blizard Institute, Queen Mary University of London, London, UKSummary: Dermal fibroblasts play a central role in the production and remodeling of dermal extracellular matrix. Their spatially distinct subpopulations exhibit different functions during skin homeostasis, wound healing, and other diseases such as fibrosis and cancer. Here, we present a protocol for detecting 3D dermal fibroblast cellular dynamics in mice without fixing the tissue using an intravital imaging technique. We describe steps for preparing, labeling, and mounting mice. We then describe detailed procedures for intravital imaging, recovery, and quantification.For complete details on the use and execution of this protocol, please refer to Rognoni et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002072Single CellMicroscopyModel Organisms
spellingShingle Toru Hiratsuka
Emanuel Rognoni
Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique
STAR Protocols
Single Cell
Microscopy
Model Organisms
title Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique
title_full Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique
title_fullStr Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique
title_full_unstemmed Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique
title_short Protocol for detecting 3D dermal fibroblast cellular dynamics in mice using an intravital imaging technique
title_sort protocol for detecting 3d dermal fibroblast cellular dynamics in mice using an intravital imaging technique
topic Single Cell
Microscopy
Model Organisms
url http://www.sciencedirect.com/science/article/pii/S2666166725002072
work_keys_str_mv AT toruhiratsuka protocolfordetecting3ddermalfibroblastcellulardynamicsinmiceusinganintravitalimagingtechnique
AT emanuelrognoni protocolfordetecting3ddermalfibroblastcellulardynamicsinmiceusinganintravitalimagingtechnique