Fig. 2
Species-mixing experiment and CTC loading in Hydro-Seq. a–d Species-mixing experiment. a Chambers with beads paired to a mouse cell (3T3 with green fluorescence) and a human cell (HEK with red fluorescence). Fluorescent imaging was applied to examine the pairing condition and verify the capture. b Histograms of the percent cross-species contamination of mixed mouse and human cells. Cells with over 90% transcripts mapped to human were labelled red and cells with over 90% transcripts mapped to mouse were labeled green. c Species-mixing analysis highlights the single-cell resolution RNA-sequencing with zero cellular cross contamination. Each red dot represents a mouse cell and each blue dot represents a human cell. d tSNE plot of single-cell expression analysis for HEK cells from two independent Hydro-Seq experiments. (Experiment 1 labeled with red color and experiment 2 labeled with cyan color) Each dot represents a cell. Cells from two experiments are evenly dispersed and do not show any clustering, indicating good reproducibility of Hydro-Seq. e–h CTC loading in Hydro-Seq. e After CTC enrichment, single-cell RNA-sequencing (scRNA-seq) of the CTCs is still challenged by the presence of many background blood cells. (Scale bar: 25 µm) f Erythrocytes flowing through the chamber during sample loading. (Scale bar: 25 µm) g During the washing cycle, the bead valve is opened to remove other blood cells through the bead capture flow channel, achieving contamination-free single CTC isolation for bead pairing. h Pairing a bead to a single CTC for scRNA-seq. Source data are provided as a Source Data file
