Isolation of Nuclei from Flash-frozen Liver Tissue for Single-cell Multiomics

The liver is a complex and heterogenous tissue responsible for carrying out many critical physiological functions, such as the maintenance of energy homeostasis metabolism xenobiotics, among others. These tasks are performed through tight coordination between hepatic parenchymal non-parenchymal cells. Additionally, various metabolic activities confined to specific areas lobule-a phenomenon called zonation. Recent advances in single-cell sequencing technologies have empowered researchers investigate heterogeneity at resolution. In tissues, including liver, harsh enzymatic and/or mechanical dissociation protocols can negatively affect viability or quality suspensions needed comprehensively characterize this organ health disease. This paper describes robust reproducible protocol isolating nuclei from frozen, archived tissues. method yields high-quality that compatible with downstream, omics approaches, single-nucleus RNA-seq, assay transposase-accessible chromatin high-throughput (ATAC-seq), well multimodal (joint RNA-seq ATAC-seq). has been successfully used isolation healthy diseased human, mouse, non-human primate frozen samples. approach allows unbiased all major cell types and, therefore, offers methodology studying