scholarly journals Droplet-based microfluidic analysis and screening of single plant cells

2017 ◽  
Author(s):  
Ziyi Yu ◽  
Christian R. Boehm ◽  
Julian M. Hibberd ◽  
Chris Abell ◽  
Jim Haseloff ◽  
...  
Keyword(s):  
High Throughput ◽  
Mammalian Cells ◽  
Plant Cells ◽  
Inducible Promoter ◽  
Microfluidic System ◽  
Marchantia Polymorpha ◽  
High Throughput Analysis ◽  
Microfluidic Analysis ◽  
On Chip ◽  
Stochastic Properties

AbstractDroplet-based microfluidics has been used to facilitate high throughput analysis of individual prokaryote and mammalian cells. However, there is a scarcity of similar workflows applicable to rapid phenotyping of plant systems. We report on-chip encapsulation and analysis of protoplasts isolated from the emergent plant model Marchantia polymorpha at processing rates of >100,000 protoplasts per hour. We use our microfluidic system to quantify the stochastic properties of a heat-inducible promoter across a population of transgenic protoplasts to demonstrate that it has the potential to assess gene expression activity in response to environmental conditions. We further demonstrate on-chip sorting of droplets containing YFP-expressing protoplasts from wild type cells using dielectrophoresis force. This work opens the door to droplet-based microfluidic analysis of plant cells for applications ranging from high-throughput characterisation of DNA parts to single-cell genomics.

scholarly journals Clarifying intact 3D tissues on a microfluidic chip for high-throughput structural analysis

2016 ◽  
Vol 113 (52) ◽  
pp. 14915-14920 ◽  
Author(s):  
Yih Yang Chen ◽  
Pamuditha N. Silva ◽  
Abdullah Muhammad Syed ◽  
Shrey Sindhwani ◽  
Jonathan V. Rocheleau ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Three Dimensional ◽  
Microfluidic System ◽  
High Throughput Drug Screening ◽  
Image Analysis Algorithm ◽  
Screening Assays ◽  
On Chip ◽  
Tissue Models

On-chip imaging of intact three-dimensional tissues within microfluidic devices is fundamentally hindered by intratissue optical scattering, which impedes their use as tissue models for high-throughput screening assays. Here, we engineered a microfluidic system that preserves and converts tissues into optically transparent structures in less than 1 d, which is 20× faster than current passive clearing approaches. Accelerated clearing was achieved because the microfluidic system enhanced the exchange of interstitial fluids by 567-fold, which increased the rate of removal of optically scattering lipid molecules from the cross-linked tissue. Our enhanced clearing process allowed us to fluorescently image and map the segregation and compartmentalization of different cells during the formation of tumor spheroids, and to track the degradation of vasculature over time within extracted murine pancreatic islets in static culture, which may have implications on the efficacy of beta-cell transplantation treatments for type 1 diabetes. We further developed an image analysis algorithm that automates the analysis of the vasculature connectivity, volume, and cellular spatial distribution of the intact tissue. Our technique allows whole tissue analysis in microfluidic systems, and has implications in the development of organ-on-a-chip systems, high-throughput drug screening devices, and in regenerative medicine.

High-throughput analysis of horse sperms’ 3D swimming patterns using computational on-chip imaging

2016 ◽  
Vol 169 ◽  
pp. 45-55 ◽  
Author(s):  
Ting-Wei Su ◽  
Inkyum Choi ◽  
Jiawen Feng ◽  
Kalvin Huang ◽  
Aydogan Ozcan
Keyword(s):  
High Throughput ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
On Chip

Pump-free multi-well-based microfluidic system for high-throughput analysis of size-control relative genes in budding yeast

2014 ◽  
Vol 6 (7) ◽  
pp. 685-693 ◽  
Author(s):  
Xianjie Kang ◽  
Lingli Jiang ◽  
Xi Chen ◽  
Haiyu Yuan ◽  
Chunxiong Luo ◽  
...  
Keyword(s):  
High Throughput ◽  
Budding Yeast ◽  
Protein Localization ◽  
Size Control ◽  
Microfluidic System ◽  
Cell Level ◽  
Protein Expression Level ◽  
Cell Cycle Protein ◽  
High Throughput Analysis ◽  
Yeast Strains

With a simple but robust well-based microfluidic device, we can high-throughput load and trace dozens of different budding yeast strains at single cell level simultaneously, providing precise cell information such as cell size, cell cycle, protein localization and protein expression level.

scholarly journals High-Efficiency and High-Throughput On-Chip Exchange of the Continuous Phase in Droplet Microfluidic Systems

2017 ◽  
Vol 22 (5) ◽  
pp. 529-535 ◽  
Author(s):  
Minkyu Kim ◽  
Chia Min Leong ◽  
Ming Pan ◽  
Lucas R. Blauch ◽  
Sindy K. Y. Tang
Keyword(s):  
High Throughput ◽  
Continuous Flow ◽  
High Efficiency ◽  
Scale Up ◽  
Continuous Phase ◽  
Microfluidic System ◽  
Droplet Surface ◽  
Generation Process ◽  
Microfluidic Systems ◽  
On Chip

This article describes an integrated platform for the on-chip exchange of the continuous phase in droplet microfluidic systems. The drops used in this work are stabilized by amphiphilic nanoparticles. For some characterizations and applications of these nanoparticle-stabilized drops, including the measurement of adsorption dynamics of nanoparticles to the droplet surface, it is necessary to change the composition of the continuous phase from that used during the droplet generation process. Thus far, no work has reported the exchange of the continuous phase for a large number (>1 million) of drops in a microfluidic system. This article describes the design and characterization of a high-efficiency and high-throughput on-chip exchanger of the continuous phase in a continuous-flow droplet microfluidic system. The efficiency of exchange was higher than 97%. The throughput was greater than 1 million drops/min, and this can be increased further by increasing the number of parallel exchangers used. Because drops are injected into the exchanger in a continuous-flow manner, the method is directly compatible with automation to further increase its reliability and potential scale-up.

scholarly journals On-Chip, Cell-Based Microarray Immunofluorescence Assay for High-Throughput Analysis of Target Proteins

2008 ◽  
Vol 80 (17) ◽  
pp. 6633-6639 ◽  
Author(s):  
Tiago G. Fernandes ◽  
Seok-Joon Kwon ◽  
Moo-Yeal Lee ◽  
Douglas S. Clark ◽  
Joaquim M. S. Cabral ◽  
...  
Keyword(s):  
High Throughput ◽  
Immunofluorescence Assay ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
Target Proteins ◽  
On Chip

scholarly journals Microfluidic system for on-chip high-throughput whole-animal sorting and screening at subcellular resolution

2007 ◽  
Vol 104 (35) ◽  
pp. 13891-13895 ◽  
Author(s):  
C. B. Rohde ◽  
F. Zeng ◽  
R. Gonzalez-Rubio ◽  
M. Angel ◽  
M. F. Yanik
Keyword(s):  
High Throughput ◽  
Microfluidic System ◽  
On Chip ◽  
Subcellular Resolution
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