{"doi":"10.1016/j.xpro.2025.103716","title":"Protocol for spatial metabolomics and isotope tracing in the mouse brain","abstract":"Mass spectrometry imaging enables high-resolution spatial chemical mapping, yet its application for dynamic analysis with tracers poses challenges. Here, we present a protocol for spatial metabolomics and isotope tracing in the mouse brain. We describe steps for tracer administration, tissue collection, and cryosectioning. We then detail procedures for matrix application, ion identification, and data analysis. This protocol delivers high-quality spatial metabolomics data and is well suited for region-specific tracing analysis in the brain. • Step-by-step workflow for spatial metabolomics and isotope tracing in the mouse brain • Guidelines for tracer administration, tissue collection, and cryosectioning • Optimized matrix deposition and MSI acquisition for high-resolution imaging • Standardized data processing for metabolite annotation and 13 C enrichment analysis Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Mass spectrometry imaging enables high-resolution spatial chemical mapping, yet its application for dynamic analysis with tracers poses challenges. Here, we present a protocol for spatial metabolomics and isotope tracing in the mouse brain. We describe steps for tracer administration, tissue collection, and cryosectioning. We then detail procedures for matrix application, ion identification, and data analysis. This protocol delivers high-quality spatial metabolomics data and is well suited for region-specific tracing analysis in the brain.","journal":"STAR Protocols","year":2025,"id":533447,"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":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9487,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":751339,"name":"Francine Yanchik-Slade","orcid":"0000-0002-3816-9990","position":1,"is_corresponding":false},{"id":1234063,"name":"Carlos Rodríguez‐Navas","orcid":"0000-0002-8499-5513","position":2,"is_corresponding":false},{"id":460221,"name":"Md Amir Hossen","orcid":null,"position":3,"is_corresponding":false},{"id":1126410,"name":"Manuel González-Rodríguez","orcid":"0000-0002-1227-2391","position":4,"is_corresponding":false},{"id":828640,"name":"Jake Vaynshteyn","orcid":"0009-0005-5596-0871","position":5,"is_corresponding":false},{"id":1415651,"name":"Shinichi Yamaguchi","orcid":null,"position":6,"is_corresponding":false},{"id":1415652,"name":"M. Nazim Boutaghou","orcid":null,"position":7,"is_corresponding":false},{"id":892316,"name":"Isaac Marin‐Valencia","orcid":null,"position":8,"is_corresponding":false},{"id":1415266,"name":"Watit Sontising","orcid":"0000-0003-3156-5832","position":0,"is_corresponding":true}],"reference_count":7,"raw_metadata":null,"created_at":"2026-07-19T02:51:32.301795Z","pmid":"40153438","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":[]}