Fig. 4.

Selection of individual hybridoma cell clones from large heterogeneous populations. Mixed populations of calcein-red/orange-stained hybridoma cells expressing 4E3 antibody and nonstained hybridoma cells expressing Elec-403 antibody in ratios of 1∶1,000 (A) and 1∶10,000 (C) were sorted. The fluorescence signals (green channel corresponding to ACE-1 activity; x axis) of the drops at the sorting junction were plotted against the droplet width (y axis) and gates for the collection of droplets with specific fluorescence intensity were set (green and red rectangles). The percentage of total droplets in each gate is indicated. The relative frequency of all events is color coded as indicated on the right. (B and D) Bright field (Top) and red/orange fluorescence (Bottom) images of the 1∶1,000 and 1∶10,000 cell dilutions before the sort and after recovery from droplets within the specific gates. The percentage of calcein-red/orange-stained hybridoma cells is indicated.