scholarly journals Double emulsion flow cytometry with high-throughput single droplet isolation and nucleic acid recovery

Lab on a Chip ◽  
2020 ◽  
Vol 20 (12) ◽  
pp. 2062-2074 ◽  
Author(s):  
Kara K. Brower ◽  
Catherine Carswell-Crumpton ◽  
Sandy Klemm ◽  
Bianca Cruz ◽  
Gaeun Kim ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nucleic Acid ◽  
High Throughput ◽  
Double Emulsion ◽  
Single Droplet ◽  
Emulsion Droplets ◽  
Acid Recovery ◽  
Emulsion Flow ◽  
Double Emulsion Droplets

We have developed a novel workflow (sdDE-FACS, s̲ingle d̲roplet D̲ouble E̲mulsion FACS) that allows robust production, screening, and sorting of single double emulsion droplets with complete nucleic acid recovery.

scholarly journals Monodisperse Water-in-Oil-in-Water (W/O/W) Double Emulsion Droplets as Uniform Compartments for High-Throughput Analysis via Flow Cytometry

Micromachines ◽  
2013 ◽  
Vol 4 (4) ◽  
pp. 402-413 ◽  
Author(s):  
Jing Yan ◽  
Wolfgang-Andreas Bauer ◽  
Martin Fischlechner ◽  
Florian Hollfelder ◽  
Clemens Kaminski ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Double Emulsion ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
Emulsion Droplets ◽  
Oil In Water ◽  
Double Emulsion Droplets

scholarly journals Optimized double emulsion flow cytometry with high-throughput single droplet isolation

2019 ◽  
Author(s):  
Kara K. Brower ◽  
Catherine Carswell-Crumpton ◽  
Sandy Klemm ◽  
Bianca Cruz ◽  
Gaeun Kim ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Double Emulsion ◽  
Droplet Microfluidics ◽  
Cell Analysis ◽  
Single Droplet ◽  
Emulsion Droplets ◽  
Droplet Sorting

Droplet microfluidics has made large impacts in diverse areas such as enzyme evolution, chemical product screening, polymer engineering, and single-cell analysis. However, while droplet reactions have become increasingly sophisticated, phenotyping droplets by a fluorescent signal and sorting them to isolate variants-of-interest remains a field-wide bottleneck. Here, we present an optimized double emulsion workflow, sdDE-FACS, that enables high-throughput phenotyping, selection, and sorting of droplets using standard flow cytometers. Using a 130 μm nozzle, we demonstrate robust post-sort recovery of intact droplets, with little to no shear-induced droplet breakage, at high sort frequency (12-14 kHz) across two industry-standard FACS instruments. We report the first quantitative plate statistics for double emulsion droplet isolation and demonstrate single droplet recovery with >70% efficiency. In addition, we establish complete downstream recovery of nucleic acids from single, sorted double emulsion droplets, an advance in droplet sorting comparable with the capabilities of single-cell FACS. This work resolves several hurdles in the field of high-throughput droplet analysis and paves the way for a variety of new droplet assays, including rare variant isolation and multiparameter single-cell analysis, marrying the full power of flow cytometry with droplet microfluidics.

scholarly journals High-Throughput Production of Micrometer Sized Double Emulsions and Microgel Capsules in Parallelized 3D Printed Microfluidic Devices

Polymers ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1887 ◽  
Author(s):  
Alexander Jans ◽  
Jonas Lölsberg ◽  
Abdolrahman Omidinia-Anarkoli ◽  
Robin Viermann ◽  
Martin Möller ◽  
...  
Keyword(s):  
Surface Modification ◽  
High Throughput ◽  
Hollow Sphere ◽  
Double Emulsion ◽  
Continuous Phase ◽  
Flow Focusing ◽  
Double Emulsions ◽  
Emulsion Droplets ◽  
3D Printed ◽  
Double Emulsion Droplets

Double emulsions are useful geometries as templates for core-shell particles, hollow sphere capsules, and for the production of biomedical delivery vehicles. In microfluidics, two approaches are currently being pursued for the preparation of microfluidic double emulsion devices. The first approach utilizes soft lithography, where many identical double-flow-focusing channel geometries are produced in a hydrophobic silicone matrix. This technique requires selective surface modification of the respective channel sections to facilitate alternating wetting conditions of the channel walls to obtain monodisperse double emulsion droplets. The second technique relies on tapered glass capillaries, which are coaxially aligned, so that double emulsions are produced after flow focusing of two co-flowing streams. This technique does not require surface modification of the capillaries, as only the continuous phase is in contact with the emulsifying orifice; however, these devices cannot be fabricated in a reproducible manner, which results in polydisperse double emulsion droplets, if these capillary devices were to be parallelized. Here, we present 3D printing as a means to generate four identical and parallelized capillary device architectures, which produce monodisperse double emulsions with droplet diameters in the range of 500 µm. We demonstrate high throughput synthesis of W/O/W and O/W/O double emulsions, without the need for time-consuming surface treatment of the 3D printed microfluidic device architecture. Finally, we show that we can apply this device platform to generate hollow sphere microgels.

scholarly journals Production of giant unilamellar vesicles and encapsulation of lyotropic nematic liquid crystals

Soft Matter ◽  
2021 ◽  
Author(s):  
Peng Bao ◽  
Daniel A. Paterson ◽  
Sally A. Peyman ◽  
J. Cliff Jones ◽  
Jonathan A. T. Sandoe ◽  
...  
Keyword(s):  
Liquid Crystals ◽  
Nematic Liquid Crystals ◽  
Double Emulsion ◽  
Lyotropic Liquid Crystals ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Emulsion Droplets ◽  
Double Emulsion Droplets

We describe a modified microfluidic method for making Giant Unilamellar Vesicles (GUVs) via water/octanol-lipid/water double emulsion droplets and encapsulation of nematic lyotropic liquid crystals (LNLCs).

scholarly journals Rapid formation of multicellular spheroids in double-emulsion droplets with controllable microenvironment

2013 ◽  
Vol 3 (1) ◽  
Author(s):  
Hon Fai Chan ◽  
Ying Zhang ◽  
Yi-Ping Ho ◽  
Ya-Ling Chiu ◽  
Youngmee Jung ◽  
...  
Keyword(s):  
Double Emulsion ◽  
Multicellular Spheroids ◽  
Emulsion Droplets ◽  
Rapid Formation ◽  
Double Emulsion Droplets

scholarly journals Fabrication of polymersomes with controllable morphologies through dewetting w/o/w double emulsion droplets

2016 ◽  
Vol 34 (4) ◽  
pp. 475-482 ◽  
Author(s):  
Wei-cai Wang ◽  
Kai Shi ◽  
Yan-xiong Pan ◽  
Chao Peng ◽  
Zi-liang Zhao ◽  
...  
Keyword(s):  
Double Emulsion ◽  
Emulsion Droplets ◽  
Double Emulsion Droplets
Sign in / Sign up

Export Citation Format

Share Document