Figure 1. Microfluidic hydrodynamic trap array for single-cell lineage tracking.

(a) Schematic representation of the hydrodynamic trap array consisting of 20 lanes of traps (inset shows an optical micrograph of a single trapped cell—scale bar, 20 μm). Independent control of upstream (P1) and downstream (P2, P3) pressures enables continuous perfusion for long-term proliferation (Supplementary Movie 2), as well as single cell release from the device (Supplementary Movie 3, Supplementary Note 1). (b) Single-cell interdivisionary time measurements for a murine leukemia cell line (L1210, n=526) and primary CD8+ T-cells isolated from C57BL/6J mice (CD8+, n=418) collected with the hydrodynamic trap array. Overlaid black bars indicate the mean interdivisionary time measured on-chip (left) and the doubling time measurements collected for bulk cultures (right) of each cell type (Supplementary Fig. 3b). (c) Overlay of lineage trees from single CD8+ T cells (n=15) and L1210 cells (n=20) established with time-lapse imaging in the hydrodynamic trap array. As a demonstration of lineage construction, one lineage for each cell type (black circles) has connecting lines indicating familial history. The first division observed in the device corresponds to time zero for each lineage with subsequent points corresponding to the second (blue circles) and third (green circles) divisions on-chip. The inset demonstrates how sister and cousin cell pairs are defined for cell lineages released for sequencing.