To sort, the system removes inactive and unwanted compounds, dumping the drops into a "bad egg" bin, and guides the others into a "keep" container. Specifically, as the drops flow through the channels they eventually encounter a junction (a two-channel fork). Left alone, the drops will naturally flow towards the path of least fluidic resistance, or the waste channel.
The device identifies the desired drops by using a laser focused on the channel before the fork to read a drop's fluorescence level. The drops with greater intensity of fluorescence (those exhibiting the highest levels of activity) are pulled towards the keep channel by the application of an electrical force, a process known as dielectrophoresis.
"Our concept was to build a miniature laboratory for performing biological experiments quickly and efficiently," explains collaborator Adam Abate, a postdoctoral fellow in applied physics at SEAS. "To do this we needed to construct microfluidic versions of common bench-top tasks, such as isolating cells in a compartment, adding reagents, and sorting the good from the bad. The challenge was to do this with microscopic drops flowing past at thousands per second."
"The sorting process is remarkably efficient and fast. By shrinking down the reaction size to 10 picoliters of volumes, we increased the sorting speed by the same amount," adds Agresti. "In our demonstration with horseradish peroxidase, we evolved and improved an already efficient enzyme by sorting through 100 million variants and choosing the best among them."
In particular, the researchers were struck by the ability to increase the efficiency of an already efficient enzyme to near its theoretical maximum, the diffusion limit, where the enzyme can produce products as quickly as a new substrate can bump into it.
|Contact: Michael Patrick Rutter|