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Droplet-based microfluidic analysis and screening of single plant cells

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...

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Detalles Bibliográficos
Autores principales: Yu, Ziyi, Boehm, Christian R., Hibberd, Julian M., Abell, Chris, Haseloff, Jim, Burgess, Steven J., Reyna-Llorens, Ivan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5933695/
https://www.ncbi.nlm.nih.gov/pubmed/29723275
http://dx.doi.org/10.1371/journal.pone.0196810
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author Yu, Ziyi
Boehm, Christian R.
Hibberd, Julian M.
Abell, Chris
Haseloff, Jim
Burgess, Steven J.
Reyna-Llorens, Ivan
author_facet Yu, Ziyi
Boehm, Christian R.
Hibberd, Julian M.
Abell, Chris
Haseloff, Jim
Burgess, Steven J.
Reyna-Llorens, Ivan
author_sort Yu, Ziyi
collection PubMed
description 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.
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spelling pubmed-59336952018-05-11 Droplet-based microfluidic analysis and screening of single plant cells Yu, Ziyi Boehm, Christian R. Hibberd, Julian M. Abell, Chris Haseloff, Jim Burgess, Steven J. Reyna-Llorens, Ivan PLoS One Research Article 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. Public Library of Science 2018-05-03 /pmc/articles/PMC5933695/ /pubmed/29723275 http://dx.doi.org/10.1371/journal.pone.0196810 Text en © 2018 Yu et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yu, Ziyi
Boehm, Christian R.
Hibberd, Julian M.
Abell, Chris
Haseloff, Jim
Burgess, Steven J.
Reyna-Llorens, Ivan
Droplet-based microfluidic analysis and screening of single plant cells
title Droplet-based microfluidic analysis and screening of single plant cells
title_full Droplet-based microfluidic analysis and screening of single plant cells
title_fullStr Droplet-based microfluidic analysis and screening of single plant cells
title_full_unstemmed Droplet-based microfluidic analysis and screening of single plant cells
title_short Droplet-based microfluidic analysis and screening of single plant cells
title_sort droplet-based microfluidic analysis and screening of single plant cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5933695/
https://www.ncbi.nlm.nih.gov/pubmed/29723275
http://dx.doi.org/10.1371/journal.pone.0196810
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