CytoSpark™ MSP Microfluidic High-throughput Screening System

The MSP High-throughput Screening System is based on the principle that fluorescent labeling and dielectrophoresis drive water-in-oil droplet sorting. A simple droplet encapsulation and sorting procedure can achieve microbial isolation and functional screening of 107 mutant or genetically engineered libraries in a single run. Unique correlated phenotypic and genotypic screening with detection of extracellular secretory proteins, membranes or intracellular markers.

Workflow
· Construction of gene mutation libraries or metagenomic libraries
Mutation libraries are constructed by random mutation, saturation mutation, etc., the metagenomic libraries can be constructed by enzyme cutting or mechanical shear plus carrier connection, and transfection to microorgansims such as E. coli, yeast, etc.
· Microdroplet generation
Using a droplet generator and accompanying chip, individual microbes are encapsulated with detection reagents in pL-level microdroplet reaction units. Separation and encapsulation can be accomplished within 10 min.
· Cell incubation
Within the microdroplet, microbes secrete enzymes that react with the substrate, or lyse and release enzymes that react with the probe to produce a fluorescent signal that can be recognized by the MSP droplet sorter.
· Droplet sorting
Utilizing dielectrophoresis and other technologies, MSP droplet sorter is capable of high-throughput detection and sorting and enrichment of positive droplets containing target strains/cells. The sorting speed can reach 200~1000 droplets/second, the enrichment efficiency is more than 95 %, and the cell yield is ≥80 %. (1 h can sort nearly one million monoclonal cells, and can sort one million to ten million clones in a single run.)
· Droplet distribution
Enables the collection of individual target microdroplets into the corresponding well plate, flexibly adapting to 96/384 well plates.

· Uniqueness
Integration of flow-sorting and plate assay for functional microbial screening with direct phenotypic and genotypic correlation, covering single or multi-parameter sorting of microbial extracellular secretion products (proteins), cell surface and intracellular markers.
· High throughput
Multi-millions of droplets which is equivalent to millions of clones sorting per run.
· High efficiency
3 to 4 orders of magnitude faster than conventional 96-well plate method.
· Time-saving
Weeks of screening by conventional methods reduced to 1-2 days.
· High sensitivity
Conventional microplates take weeks to detect secreted proteins per individual microbe, whereas the MSP system's nL-grade water-in-oil system encapsulates single cells and detects secreted proteins in a few hours.
· Low cost
Reduced reagent usage, reagent consumption down to one millionth of conventional methods.