Fig. 4

Flow cell device for single-cell RNA-Seq. a Graphical representation of our five-lane microwell array flow cell device for single-cell RNA-Seq. b Schematic of on-chip steps for single-cell RNA-Seq. After depositing cells, barcoded capture beads (barcode sequences represented as different colors), and sealing as in Fig. 2a, single-cell lysates (green) are trapped in individual microwells and mRNA hybridizes to the barcoded capture beads. The device is unsealed and rapidly washed by flow before on-chip, solid-phase reverse transcription and second-strand synthesis followed by elution and pre-amplification of the pooled library by in vitro transcription. c Montage of fluorescence images from part of one lane of the device in (a) showing beads (red) and cells (blue) loaded in the array. Note that this image was acquired following cell lysis while the device is sealed, and so the blue live stain fills the entire volume of the corresponding microwell and is confined to the microwell by sealing