Protocol for integrating tissue clearing and light-sheet imaging to analyze cancer initiation in mosaic analysis with double markers mouse models.

STAR Protoc

Department of Microbiology, Immunology, and Cancer Biology, University of Virginia Health System, Charlottesville, VA 22908, USA; University of Virginia Cancer Center, University of Virginia Health System, Charlottesville, VA 22903, USA. Electronic address:

Published: June 2024

Mosaic analysis with double markers (MADM) mouse models closely mimic the clonal origin of human cancers by generating sporadic, GFP-labeled cancer-initiating cells. Traditional clonal analysis pipelines are labor intensive, hindering throughput and disrupting the 3D architecture. Here, we present a protocol that integrates tissue clearing and light-sheet imaging to analyze pre-malignant clones in whole-mount MADM-labeled tissues. We describe steps for generating mosaic-labeled cancer mouse models, tissue harvesting, fixation, and clearing. We then detail procedures for light-sheet imaging and clonal size analysis. For complete details on the use and execution of this protocol, please refer to Zeng et al..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11152722PMC
http://dx.doi.org/10.1016/j.xpro.2024.103092DOI Listing

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