A bulk sub-femtoliter in vitro compartmentalization system using super-fine electrosprays

The extreme miniaturization of biological and chemical assays in aqueous-droplet compartments enables spatiotemporal control for large-scale parallel experimentation and can thus permit new capabilities for “digitizing” directed molecular evolution methodologies. We report a remarkably facile bulk m...

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Bibliographic Details
Published in:Scientific reports 2016-05, Vol.6 (1), p.26257-26257, Article 26257
Main Authors: Sharma, Bineet, Takamura, Yuzuru, Shimoda, Tatsuya, Biyani, Manish
Format: Article
Language:English
Subjects:
631/1647/2163
631/61/350/1057
Chemical reactions
Digitizing
Encapsulation
Evolution
Gene expression
Homogenization
Humanities and Social Sciences
Molecular evolution
multidisciplinary
Proteins
Science
Science (multidisciplinary)
Surfactants
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Summary:The extreme miniaturization of biological and chemical assays in aqueous-droplet compartments enables spatiotemporal control for large-scale parallel experimentation and can thus permit new capabilities for “digitizing” directed molecular evolution methodologies. We report a remarkably facile bulk method to generate mega-scale monodisperse sub-femtoliter aqueous droplets by electrospray, using a prototype head with super-fine inkjet technology. Moreover, the electrostatic inkjet nozzle that injects the aqueous phase when immersed within an immiscible phase (an optimized oil/surfactant mixture) has the advantage of generating cell-like sub-femtoliter compartments for biomolecule encapsulation and successive biological and chemical reactions. Sub-femtoliter droplets of both liquid (water-in-oil, volumes ranging from 0.2 to 6.4 fL) and gel bead (agarose-in-oil, volume ranging from 0.3 to 15.6 fL) compartments with average sizes of 1.3 μm and 1.5 μm, respectively, were successfully generated using an inkjet nozzle at a speed of more than 10 5 droplets per second. We demonstrated the applicability of this system by synthesizing fluorescent proteins using a cell-free expression system inside electrosprayed sub-femtoliter droplets at an accelerated rate, thereby extending the utility of in vitro compartmentalization with improved analytical performance for a top-down artificial cellular system.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep26257