Droplet-based microfluidic analysis and screening of single plant cells.
Burgess, Steven J
Public Library of Science (PLoS)
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Yu, Z., Boehm, C., Hibberd, J., Abell, C., Haseloff, J., Burgess, S. J., & Reyna-Llorens, I. (2018). Droplet-based microfluidic analysis and screening of single plant cells.. PloS one, 13 (5), e0196810. https://doi.org/10.1371/journal.pone.0196810
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. 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.
Protoplasts, Plants, Genetically Modified, Marchantia, Bacterial Proteins, Luminescent Proteins, Microscopy, Fluorescence, Microfluidic Analytical Techniques, Cell Separation, Stochastic Processes, Equipment Design, Drug Compounding, Genomics, Gene Expression Regulation, Plant, Transformation, Genetic, Genes, Reporter, Hot Temperature, Promoter Regions, Genetic, Lab-On-A-Chip Devices, High-Throughput Screening Assays, Single-Cell Analysis, Agrobacterium tumefaciens
BBSRC and EPSRC OpenPlant BB/L014130/1
External DOI: https://doi.org/10.1371/journal.pone.0196810
This record's URL: https://www.repository.cam.ac.uk/handle/1810/278891
Attribution 4.0 International
Licence URL: http://creativecommons.org/licenses/by/4.0/
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