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Droplet-based microfluidic analysis and screening of single plant cells.

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Yu, Ziyi 
Boehm, Christian R 
Hibberd, Julian M 
Abell, Chris 
Haseloff, Jim 


Droplet-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 where phenotyping analyses typically are time-consuming and low-throughput. We report on-chip encapsulation and analysis of protoplasts isolated from the emergent plant model Marchantia polymorpha at processing rates of >100,000 cells 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 its potential for assessing 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 to selection of rare plant phenotypes.



Agrobacterium tumefaciens, Bacterial Proteins, Cell Separation, Drug Compounding, Equipment Design, Gene Expression Regulation, Plant, Genes, Reporter, Genomics, High-Throughput Screening Assays, Hot Temperature, Lab-On-A-Chip Devices, Luminescent Proteins, Marchantia, Microfluidic Analytical Techniques, Microscopy, Fluorescence, Plants, Genetically Modified, Promoter Regions, Genetic, Protoplasts, Single-Cell Analysis, Stochastic Processes, Transformation, Genetic

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PLoS One

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Public Library of Science (PLoS)
Biotechnology and Biological Sciences Research Council (BB/L014130/1)
BBSRC and EPSRC OpenPlant BB/L014130/1