Application note: bacteria and yeast encapsulation method in small double emulsions

In recent years, the microencapsulation of Bacteria and Yeast has expanded due to the advantages and novel information provided by these methods. In particular, droplet microfluidics has become a popular technique for bacterial and yeast encapsulation due to its performance, efficiency, and precision. This method has been used in various areas of microbiology, including pathogen detection and identification, antibiotic susceptibility testing, microbial physiology studies, and biotechnological applications.

Bacteria and Yeast encapsulation is a crucial technique in the investigation of microorganisms that play a vital role in the ecosystem and have a potent metabolic capacity for producing biopharmaceuticals and recombinant proteins.

To perform this encapsulation method, double water-in-oil-in-water (W/O/W) emulsions containing the microorganisms of interest are generated. Unlike single water-in-oil (W/O) emulsions, double emulsions provide an aqueous carrier fluid, making them compatible with most flow cytometry and cell sorting setups. They also protect sensitive biological structures from the sheath fluid, preserve the link between microorganisms and the substances they secrete, and allow for in vitro analysis of biomolecules.

Droplet-based microfluidic techniques, particularly for the microencapsulation of Bacteria and Yeast, offer superior control over droplet generation, producing highly stable and monodisperse double emulsions. To be compatible with cell sorting and analysis devices, the droplets must be small (< 60µm) in diameter and large enough to encapsulate the variants of interest.

Therefore, in this application note, we demonstrate an easy-to-use and reliable workflow for encapsulating the yeast strain S. Cerevisiae CEN.PK 113-7D and the bacterial strain L. cremoris MG1363_GFP into highly monodisperse double emulsions with a diameter of 42 μm, suitable for high-throughput screening and sorting. The workflow uses the Cell Encapsulation Platform, consisting of Fluigent’s fluid handling system and Secoya’s emulsification technology, the RayDrop®.

This application note was created in collaboration with Fluigent and TU Delft.

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