{"doi":"10.21769/bioprotoc.4601","title":"Isolation of Nuclei from Human Snap-frozen Liver Tissue for Single-nucleus RNA Sequencing","abstract":"Single-nucleus RNA sequencing (snRNA-seq) provides a powerful tool for studying cell type composition in heterogenous tissues. The liver is a vital organ composed of a diverse set of cell types; thus, single-cell technologies could greatly facilitate the deconvolution of liver tissue composition and various downstream omics analyses at the cell-type level. Applying single-cell technologies to fresh liver biopsies can, however, be very challenging, and snRNA-seq of snap-frozen liver biopsies requires some optimization given the high nucleic acid content of the solid liver tissue. Therefore, an optimized protocol for snRNA-seq specifically targeted for the use of frozen liver samples is needed to improve our understanding of human liver gene expression at the cell-type resolution. We present a protocol for performing nuclei isolation from snap-frozen liver tissues, as well as guidance on the application of snRNA-seq. We also provide guidance on optimizing the protocol to different tissue and sample types.","journal":"BIO-PROTOCOL","year":2023,"id":380583,"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":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9352,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":278348,"name":"Jihane N. Benhammou","orcid":"0000-0003-2442-5145","position":1,"is_corresponding":false},{"id":580546,"name":"Shuyun Rao","orcid":"0000-0002-1073-6682","position":2,"is_corresponding":false},{"id":420257,"name":"Lopa Mishra","orcid":"0000-0002-6850-0808","position":3,"is_corresponding":false},{"id":278349,"name":"Joseph R. Pisegna","orcid":"0000-0002-5442-2474","position":4,"is_corresponding":false},{"id":49873,"name":"Päivi Pajukanta","orcid":"0000-0002-6423-8056","position":5,"is_corresponding":false},{"id":49869,"name":"Marcus Alvarez","orcid":"0000-0002-6369-5028","position":0,"is_corresponding":true}],"reference_count":22,"raw_metadata":null,"created_at":"2026-07-19T01:17:04.905707Z","pmid":"36874905","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":[]}