scholarly journals Microdroplet-based system for culturing of environmental microorganisms using FNAP-sort

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kanako Saito ◽  
Yuri Ota ◽  
Dieter M. Tourlousse ◽  
Satoko Matsukura ◽  
Hirotsugu Fujitani ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
High Purity ◽  
Droplet Microfluidics ◽  
Individual Growth ◽  
Proof Of Concept ◽  
Widespread Adoption ◽  
Technical Challenges ◽  
On Chip ◽  
Uncultured Microorganisms ◽  
Environmental Microbes

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.

scholarly journals Droplet microfluidics: from proof-of-concept to real-world utility?

2019 ◽  
Vol 55 (67) ◽  
pp. 9895-9903 ◽  
Author(s):  
Akkapol Suea-Ngam ◽  
Philip D. Howes ◽  
Monpichar Srisa-Art ◽  
Andrew J. deMello
Keyword(s):  
Real World ◽  
High Speed ◽  
Droplet Microfluidics ◽  
Proof Of Concept ◽  
Conventional Instrumentation ◽  
Practical Tool ◽  
Tool Set

Droplet microfluidics constitutes a diverse and practical tool set that enables chemical and biological experiments to be performed at high speed and with enhanced efficiency when compared to conventional instrumentation.

scholarly journals Proof-of-concept demonstration of free-form optics enhanced confocal Raman spectroscopy in combination with optofluidic lab-on-chip

2016 ◽  
Author(s):  
Qing Liu ◽  
Diane De Coster ◽  
Damien Loterie ◽  
Jürgen Van Erps ◽  
Michael Vervaeke ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Free Form ◽  
Proof Of Concept ◽  
Confocal Raman Spectroscopy ◽  
Lab On Chip ◽  
Confocal Raman ◽  
On Chip

scholarly journals Progress and Future Prospectives in Skin-on-Chip Development with Emphasis on the use of Different Cell Types and Technical Challenges

2017 ◽  
Vol 13 (3) ◽  
pp. 418-429 ◽  
Author(s):  
Lenie J. van den Broek ◽  
Lambert I. J. C. Bergers ◽  
Christianne M. A. Reijnders ◽  
Susan Gibbs
Keyword(s):  
Cell Types ◽  
Technical Challenges ◽  
On Chip ◽  
Different Cell Types

scholarly journals Dual sequentially addressable dielectrophoretic array for high-throughput, scalable, multiplexed droplet sorting

2021 ◽  
Vol 25 (4) ◽  
Author(s):  
Akihiro Isozaki ◽  
Dunhou Huang ◽  
Yuta Nakagawa ◽  
Keisuke Goda
Keyword(s):  
High Throughput ◽  
Cell Biology ◽  
Digital Pcr ◽  
Industrial Applications ◽  
Droplet Microfluidics ◽  
Diverse Range ◽  
Droplet Sorting ◽  
On Chip ◽  
Single Droplets

AbstractDroplet microfluidics is a powerful tool for a diverse range of biomedical and industrial applications such as single-cell biology, synthetic biology, digital PCR, biosafety monitoring, drug screening, and food, feed, and cosmetic industries. As an integral part of droplet microfluidics, on-chip multiplexed droplet sorting has recently gained enthusiasm, since it enables real-time sorting of single droplets containing cells with different phenotypes into multiple bins. However, conventional sorting methods are limited in throughput and scalability. Here, we present high-throughput, scalable, multiplexed droplet sorting by employing a pair of sequentially addressable dielectrophoretic arrays (SADAs) across a microchannel on a microfluidic chip. A SADA is an on-chip array of electrodes, each of which is sequentially activated and deactivated in synchronization to the position and speed of a flowing droplet of interest. The dual-SADA (dSADA) structure enables high-throughput deflection of droplets in multiple directions in a well-controlled manner. For proof-of-concept demonstration and characterization of the dSADA, we performed fluorescence-activated droplet sorting (FADS) with a 3-way dSADA at a high throughput of 2450 droplets/s. Furthermore, to show the scalability of the dSADA, we also performed FADS with a 5-way dSADA at a high throughput of 473 droplets/s.

scholarly journals Kinetics of protein-assisted nucleic acid interconversion monitored by transient time resolved fluorescence in microfluidic droplets

2021 ◽  
Author(s):  
Natalia Grytsyk ◽  
Damien Cianfarani ◽  
Olivier Crégut ◽  
Ludovic Richert ◽  
Christian Boudier ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Single Molecule ◽  
Translational Regulation ◽  
Weighted Average ◽  
Resonance Energy ◽  
Droplet Microfluidics ◽  
Time Resolved Fluorescence ◽  
Transient Time ◽  
Time Resolved ◽  
Hiv 1

Abstract Interconversions between nucleic acid structures play an important role in transcriptional and translational regulation and also in repair and recombination. These interconversions are frequently promoted by nucleic acid chaperone proteins. To monitor their kinetics, Förster resonance energy transfer (FRET) is widely exploited using ensemble fluorescence intensity measurements in pre-steady-state stopped-flow experiments. Such experiments only provide a weighted average of the emission of all species in solution and consume large quantities of materials. Herein, we lift these limitations by combining time-resolved fluorescence (TRF) with droplet microfluidics (DmF). We validate the innovative TRF-DmF approach by investigating the well characterized annealing of the HIV-1 (+)/(–) Primer Binding Sequences (PBS) promoted by a HIV-1 nucleocapsid peptide. Upon rapid mixing of the FRET-labelled (–)PBS with its complementary (+)PBS sequence inside microdroplets, the TRF-DmF set-up enables resolving the time evolution of sub-populations of reacting species and reveals an early intermediate with a ∼50 ps donor fluorescence lifetime never identified so far. TRF-DmF also favorably compares with single molecule experiments, as it offers an accurate control of concentrations with no upper limit, no need to graft one partner on a surface and no photobleaching issues.

scholarly journals KLu(WO4)2/SiO2 Tapered Waveguide Platform for Sensing Applications

Micromachines ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 454 ◽  
Author(s):  
Medina ◽  
Rüter ◽  
Pujol ◽  
Kip ◽  
Masons ◽  
...  
Keyword(s):  
Inductively Coupled Plasma ◽  
Proof Of Concept ◽  
Inductively Coupled ◽  
Sensing Applications ◽  
Tapered Waveguide ◽  
Processing Step ◽  
Index Contrast ◽  
High Index Contrast ◽  
On Chip ◽  
Tapered Waveguides

This paper provides a generic way to fabricate a high-index contrast tapered waveguide platform based on dielectric crystal bonded on glass for sensing applications. As a specific example, KLu(WO4)2 crystal on a glass platform is made by means of a three-technique combination. The methodology used is on-chip bonding, taper cutting with an ultra-precise dicing saw machine and inductively coupled plasma-reactive ion etching (ICP-RIE) as a post-processing step. The high quality tapered waveguides obtained show low surface roughness (25 nm at the top of the taper region), exhibiting propagation losses estimated to be about 3 dB/cm at 3.5 m wavelength. A proof-of-concept with crystal-on-glass tapered waveguides was realized and used for chemical sensing.

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