Fig. 1
Workflow for single-nuclei isolation starting with low-input cryopreserved tissues. Outline of the main steps of the process for isolating and preparing single-nuclei for transcriptomics analyses starting with low-input cryopreserved human tissues. Module 1—Homogenization: Tissue samples are mechanically homogenized to release single nuclei while preserving nuclear integrity using a Dounce homogenizer. Module 2—Nuclei Sorting: The homogenized mixture is subjected to fluorescence-activated cell sorting (FACS) to isolate and purify the nuclei based on integrity and size. Module 3—Quality Control: Isolated nuclei undergo quality control assessment with microscopic examination to confirm their integrity and purity. Module 4—Tagging and Sequencing: Nuclei are tagged with unique molecular identifiers (UMIs) and barcoded using the Chromium platform. The tagged nuclei are then subjected to library preparation and high-throughput sequencing followed by data analysis.
