Automated processing of solid tissues into single cells or nuclei for genomics and cell biology applications with the Singulator™ 100 system

Single-cell sequencing is revealing the next level of complexity of biological systems. The ability to measure the heterogeneity of single-cell gene expression and chromatin structure cell by cell has led to the discovery of new cell types and expanded our understanding of the diverse regulatory pathways and states of individual cells. Single-cell sequencing workflows for multiple applications require reproducible generation of high-quality single-cell or nuclei suspensions. \nS2 Genomics has developed and now commercialized the patent-pending Singulator™ 100 System to automate the dissociation of solid tissues into single cell or nuclei suspensions. The operation of the commercial Singulator 100 System for the dissociation of mouse and human solid tissues will be described, along with new, sample cartridges for improved processing of samples <10 mg. Validation data will be presented for processing 5 to 400 mg samples of fresh mouse and human solid tissues, including matched sets of normal and cancerous patient samples, into single-cell suspensions with high viabilities and yields. In addition, data of processing fresh or frozen mouse and human tissue samples into high-quality nuclei in <6 min will be presented. The Singulator-prepared samples have been analyzed by single-cell RNA sequencing (scRNA-seq), single-nucleus RNA sequencing (snRNA-seq), and assay for transposase-accessible chromatin using sequencing (ATAC-seq) with a nanodroplet single-cell library preparation system. The best methods for downstream processing, including additional sample purification and use of RNAse inhibitors will be discussed. In addition, data will be presented on the use of the Singulator to isolate specific cell types from tumor samples will be shown. Supported in part by National Institutes of Health award number R41 HG010129-01 to Dr. Jovanovich.