{"doi":"10.1016/j.xpro.2024.103303","title":"Protocol to characterize mouse intestinal epithelial cell lineage using Opal multiplex immunofluorescence","abstract":"Applying Opal multiplex immunofluorescence (OMI) to characterize intestinal tissues of genetically engineered mouse models provides an excellent tool for studying complex processes. However, detecting appropriate signals from multiple target molecules is challenging. Here, we present a protocol to characterize mouse intestinal epithelial cell lineage using OMI. We describe steps for processing small intestine and colonic mouse tissues and designing and optimizing panels for OMI in mouse intestinal tissues. We then detail procedures for performing a quantitative evaluation of acquired images. For complete details on the use and execution of this protocol, please refer to Kinoshita et al. 1 • Steps for small intestine and colon processing and fixation • Multiplex imaging to delineate intestinal regeneration and tumor development • Steps for cell detection and image quantification Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Applying Opal multiplex immunofluorescence (OMI) to characterize intestinal tissues of genetically engineered mouse models provides an excellent tool for studying complex processes. However, detecting appropriate signals from multiple target molecules is challenging. Here, we present a protocol to characterize mouse intestinal epithelial cell lineage using OMI. We describe steps for processing small intestine and colonic mouse tissues and designing and optimizing panels for OMI in mouse intestinal tissues. We then detail procedures for performing a quantitative evaluation of acquired images.","journal":"STAR Protocols","year":2024,"id":503208,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9499,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":267707,"name":"Hiroto Kinoshita","orcid":"0000-0002-8179-6017","position":1,"is_corresponding":false},{"id":382238,"name":"Tania Cid-Diaz","orcid":null,"position":2,"is_corresponding":false},{"id":257674,"name":"Ángeles Durán","orcid":"0000-0001-9576-0587","position":3,"is_corresponding":false},{"id":856206,"name":"Marta Osrodek","orcid":"0000-0002-2575-121X","position":4,"is_corresponding":false},{"id":233462,"name":"María T. Diaz‐Meco","orcid":"0000-0003-0147-0998","position":5,"is_corresponding":false},{"id":233463,"name":"Jorge Moscat","orcid":"0000-0002-9442-3834","position":6,"is_corresponding":false},{"id":267706,"name":"Anxo Martínez-Ordóñez","orcid":"0000-0001-9649-2967","position":0,"is_corresponding":true}],"reference_count":3,"raw_metadata":null,"created_at":"2026-07-19T02:10:27.781502Z","pmid":"39264806","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}