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A bulk sub-femtoliter in vitro compartmentalization system using super-fine electrosprays
Bineet Sharma1, Yuzuru Takamura1,2, Tatsuya Shimoda1,2
1Department of Bioscience and Biotechnology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan.
Scientific Reports
|May 21, 2016
Summary
Researchers developed a bulk electrospray method to create millions of tiny, cell-like compartments for biological experiments. This technique accelerates molecular evolution and enhances artificial cellular systems.
Area of Science:
- Biotechnology
- Chemical Engineering
- Molecular Biology
Background:
- Miniaturized biological and chemical assays in droplet compartments offer spatiotemporal control for large-scale parallel experimentation.
- This enables new capabilities for digitizing directed molecular evolution methodologies.
Purpose of the Study:
- To report a facile bulk method for generating mega-scale monodisperse sub-femtoliter aqueous droplets using electrospray.
- To demonstrate the utility of these droplets as cell-like compartments for biomolecule encapsulation and reactions.
Main Methods:
- Utilized a prototype electrospray head with super-fine inkjet technology.
- Employed an electrostatic inkjet nozzle immersed in an immiscible oil/surfactant mixture to generate aqueous droplets.
- Generated both liquid (water-in-oil) and gel bead (agarose-in-oil) compartments with volumes ranging from sub-femtoliters to femtoliters.
Main Results:
- Successfully generated monodisperse sub-femtoliter droplets (0.2–6.4 fL) and gel beads (0.3–15.6 fL) at a rate exceeding 10^5 droplets per second.
- Achieved average droplet and gel bead sizes of 1.3 μm and 1.5 μm, respectively.
- Demonstrated accelerated synthesis of fluorescent proteins using a cell-free expression system within these droplets.
Conclusions:
- The developed electrospray method provides a high-throughput, scalable approach for creating sub-femtoliter compartments.
- This technique enhances in vitro compartmentalization for artificial cellular systems, improving analytical performance.
- The system facilitates rapid molecular evolution and biomolecular synthesis in cell-like microenvironments.
