Droplet-based microfluidic analysis and screening of single plant cells

• Published in: PloS one Vol. 13; no. 5; p. e0196810
• Main Authors: Yu, Ziyi, Boehm, Christian R, Hibberd, Julian M, Abell, Chris, Haseloff, Jim, Burgess, Steven J, Reyna-Llorens, Ivan
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 03.05.2018; Public Library of Science (PLoS)
• Subjects: Agrobacterium; Agrobacterium tumefaciens - genetics; Analysis; Bacterial Proteins - analysis; Bacterial Proteins - genetics; Biology and Life Sciences; Cell Separation - instrumentation; Cell Separation - methods; Cells (Biology); Deoxyribonucleic acid; Dielectrophoresis; DNA; Drug Compounding; Engineering and Technology; Environmental conditions; Equipment Design; Gene expression; Gene Expression Regulation, Plant; Genes, Reporter; Genetic engineering; Genomes; Genomics - methods; High-Throughput Screening Assays - instrumentation; High-Throughput Screening Assays - methods; Hot Temperature; Lab-On-A-Chip Devices; Laboratories; Luminescent Proteins - analysis; Luminescent Proteins - genetics; Mammalian cells; Marchantia - chemistry; Marchantia - cytology; Marchantia - genetics; Marchantia polymorpha; Medical screening; Medicine and Health Sciences; Microfluidic Analytical Techniques - methods; Microfluidics; Microscopy, Fluorescence; Phenotypes; Phenotyping; Physical Sciences; Physiology; Plant biology; Plant cells; Plant sciences; Plants, Genetically Modified; Prokaryotes; Promoter Regions, Genetic; Protoplasts; Protoplasts - chemistry; Single-Cell Analysis - instrumentation; Single-Cell Analysis - methods; Stochastic Processes; Stochasticity; Synthetic biology; Transformation, Genetic; United Kingdom > UK; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 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 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.