Before revision: Method 250px | In vitro compartmentalization : selection cycle Comparison of in vivo cell compartmentalisation (left) versus in vitro compartmentalization (right) using double emulsion droplets and fluorescent sorting.1) In an in vivo system, a library of genes is transformed into bacteria to produce the protein variants and dispersed into a water‐in‐oil (w/o) emulsion aiming to have each droplet contain maximum one cell.2) In an in vitro system, no cells are used and in stead protein variants are produced via in vitro transcription/translation aiming to have one gene per droplet.3,4) In both cases, droplets are then resuspended in water to make a water‐in‐oil‐in‐water (w/o/w) emulsion, have a fluorogenic substrate added, and passed in front of the laser and detector in a fluorescence activated cell sorter.
After revision: Method 400x400px | Comparison of in vivo cell compartmentalisation (left) versus in vitro compartmentalization (right) using double emulsion droplets and fluorescent sorting.1) In an in vivo system, a library of genes is transformed into bacteria to produce the protein variants and dispersed into a water‐in‐oil (w/o) emulsion aiming to have each droplet contain maximum one cell.2) In an in vitro system, no cells are used and in stead protein variants are produced via in vitro transcription/translation aiming to have one gene per droplet.3,4) In both cases, droplets are then resuspended in water to make a water‐in‐oil‐in‐water (w/o/w) emulsion, have a fluorogenic substrate added, and passed in front of the laser and detector in a fluorescence activated cell sorter.