Fig. 2

Main steps in the scRNA-seq workflow. First, the tissue of interest is dissociated to make a single-cell suspension. Single cells are then harvested for scRNA- seq analysis. Magnetic activated cell sorting relies on the immunoreactivity of cell specific antigens with magnetic beads. A fluorescence activated cell sorting platform then selects individual cells with heterogeneous tissue by detecting fluorescent labelled signals. The cells can be isolated using a variety of parameters. Smart-seq2 and CEL-Seq2 are performed in 96 or 384-well plates after sorting, while droplet systems (e.g. 10X Chromium and Drop-Seq) couple cells with barcoded beads containing a unique molecule identifier (UMI) and primers that form water in-oil droplets via a continuous oil flow. Reverse transcription and cDNA amplification are performed by PCR in Smart-Seq and 10X Chromium and by in vitro transcription (IVT) in CEL-Seq2