Figure 2. Characterization of experimental parameters for cell loading and maintenance.

A. The dependence of cell loading efficiency on the flow rate during cell loading. Cells with OD₆₀₀ = 0.25 were loaded to separate modules of the multiplexed microfluidic platform, with each module experiencing a different culture flow rate for 5 minutes: 10μL/min, 30μL/min, or 50μL/min. The loading efficiency was calculated as the percentage of fillable units out of 1,120 replicator units investigated. A replicator unit was defined as fillable if it was not initially occupied by a cell, but it had a nearby cell touching one of its three gaps. For each flow rate, a rectangular region consisting of 1,120 replicator units provided the total number of replicator units investigated. B. The dependence of cell loading efficiency on the cell density during cell loading. Using the cell loading flow rate of 30μL/min for 5 minutes, three different cell densities (OD₆₀₀ = 0.1, 0.25, 0.4) were characterized. As described in (A), the cell loading efficiency was calculated for each case. C. The dependence of cell loading efficiency on loading duration. Using the cell loading flow rate of 30μL/min and cell density of OD₆₀₀ = 0.25, three different loading durations (3min, 5min, 10min) were characterized. As described in (A), the cell loading efficiency was calculated for each case. D. The effect of the media flush rate on the percentage of mother cells removed from their traps. The reduction in the percentage of the initially-trapped cells was separately measured for three different media flush rates: 10μL/min, 30μL/min, and 50μL/min. For each case, the media flush was applied for 5 minutes and the number of initially-trapped cells served as the denominator in calculating the percent reduction. In all panels, error bars are SEM (N=2).