Fig. 2.

Sorting of 4E3 hybridoma cells mixed with a 75-fold excess of unrelated control cells. A population of calcein-red/orange-stained hybridoma cells expressing 4E3 antibody and nonstained hybridoma cells expressing Elec-403 antibody was mixed in a 1∶75 ratio and 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 relative frequency of all events is color coded as indicated on the right. Bright field (Top) and red/orange fluorescence (Bottom) images shows the nonsorted cell population as well as cells recovered from droplets within the specific gates. The percentage of calcein-red/orange-stained hybridoma cells is indicated.