Ultrahigh-Throughput Screening of Metagenomic Libraries Using Droplet Microfluidics

2021 ◽  
pp. 19-32
Author(s):  
Davide Agostino Cecchini ◽  
Mercedes Sánchez-Costa ◽  
Alejandro H. Orrego ◽  
Jesús Fernández-Lucas ◽  
Aurelio Hidalgo
Micromachines ◽  
2017 ◽  
Vol 8 (8) ◽  
pp. 230 ◽  
Author(s):  
Elias Theodorou ◽  
Randall Scanga ◽  
Mariusz Twardowski ◽  
Michael Snyder ◽  
Eric Brouzes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kanako Saito ◽  
Yuri Ota ◽  
Dieter M. Tourlousse ◽  
Satoko Matsukura ◽  
Hirotsugu Fujitani ◽  
...  

AbstractDroplet microfluidics has emerged as a powerful technology for improving the culturing efficiency of environmental microorganisms. However, its widespread adoption has been limited due to considerable technical challenges, especially related to identification and manipulation of individual growth-positive droplets. Here, we combined microfluidic droplet technology with on-chip “fluorescent nucleic acid probe in droplets for bacterial sorting” (FNAP-sort) for recovery of growth-positive droplets and droplet microdispensing to establish an end-to-end workflow for isolation and culturing of environmental microbes. As a proof-of-concept, we demonstrate the ability of our technique to yield high-purity cultures of rare microorganisms from a representative complex environmental microbiome. As our system employs off-the-shelf commercially available equipment, we believe that it can be readily adopted by others and may thus find widespread use toward culturing the high proportion of as-of-yet uncultured microorganisms in different biomes.


Micromachines ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 272
Author(s):  
Doojin Lee ◽  
Amy Q. Shen

Droplet microfluidics provides a versatile tool for measuring interfacial tensions between two immiscible fluids owing to its abilities of fast response, enhanced throughput, portability and easy manipulations of fluid compositions, comparing to conventional techniques. Purely homogeneous extension in the microfluidic device is desirable to measure the interfacial tension because the flow field enables symmetric droplet deformation along the outflow direction. To do so, we designed a microfluidic device consisting of a droplet production region to first generate emulsion droplets at a flow-focusing area. The droplets are then trapped at a stagnation point in the cross junction area, subsequently being stretched along the outflow direction under the extensional flow. These droplets in the device are either confined or unconfined in the channel walls depending on the channel height, which yields different droplet deformations. To calculate the interfacial tension for confined and unconfined droplet cases, quasi-static 2D Darcy approximation model and quasi-static 3D small deformation model are used. For the confined droplet case under the extensional flow, an effective viscosity of the two immiscible fluids, accounting for the viscosity ratio of continuous and dispersed phases, captures the droplet deformation well. However, the 2D model is limited to the case where the droplet is confined in the channel walls and deforms two-dimensionally. For the unconfined droplet case, the 3D model provides more robust estimates than the 2D model. We demonstrate that both 2D and 3D models provide good interfacial tension measurements under quasi-static extensional flows in comparison with the conventional pendant drop method.


2021 ◽  
pp. 2000123
Author(s):  
Pantelitsa Dimitriou ◽  
Jin Li ◽  
Giusy Tornillo ◽  
Thomas McCloy ◽  
David Barrow

Author(s):  
Jianhua Yang ◽  
Ran Tu ◽  
Huiling Yuan ◽  
Qinhong Wang ◽  
Leilei Zhu

Micromachines ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 662
Author(s):  
Nikita A. Filatov ◽  
Anatoly A. Evstrapov ◽  
Anton S. Bukatin

Droplet microfluidics is an extremely useful and powerful tool for industrial, environmental, and biotechnological applications, due to advantages such as the small volume of reagents required, ultrahigh-throughput, precise control, and independent manipulations of each droplet. For the generation of monodisperse water-in-oil droplets, usually T-junction and flow-focusing microfluidic devices connected to syringe pumps or pressure controllers are used. Here, we investigated droplet-generation regimes in a flow-focusing microfluidic device induced by the negative pressure in the outlet reservoir, generated by a low-cost mini diaphragm vacuum pump. During the study, we compared two ways of adjusting the negative pressure using a compact electro-pneumatic regulator and a manual airflow control valve. The results showed that both types of regulators are suitable for the stable generation of monodisperse droplets for at least 4 h, with variations in diameter less than 1 µm. Droplet diameters at high levels of negative pressure were mainly determined by the hydrodynamic resistances of the inlet microchannels, although the absolute pressure value defined the generation frequency; however, the electro-pneumatic regulator is preferable and convenient for the accurate control of the pressure by an external electric signal, providing more stable pressure, and a wide range of droplet diameters and generation frequencies. The method of droplet generation suggested here is a simple, stable, reliable, and portable way of high-throughput production of relatively large volumes of monodisperse emulsions for biomedical applications.


2018 ◽  
Vol 112 (6) ◽  
pp. 063701 ◽  
Author(s):  
Anna Fornell ◽  
Kevin Cushing ◽  
Johan Nilsson ◽  
Maria Tenje

RSC Advances ◽  
2013 ◽  
Vol 3 (46) ◽  
pp. 24423 ◽  
Author(s):  
S. N. Beesabathuni ◽  
J. G. Stockham ◽  
J. H. Kim ◽  
H. B. Lee ◽  
J. H. Chung ◽  
...  

Author(s):  
Gregor Tegl ◽  
Peter Rahfeld ◽  
Katharina Ostmann ◽  
John Hanson ◽  
Stephen G. Withers

Screening of a human gut metagenomic library yielded a set of novel beta-N-acetyl-glucosaminidases. Mutation of active site residues yielded a thioglycoligase that efficiently S-GlcNAcylates protein targets as well as various thiosugar acceptors.


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