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Droplet-based microfluidic screening and sorting of microalgal populations for strain engineering applications.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Leontidou, Tina 
Young, Rosanna EB 
Vonlanthen, Sofie E 

Abstract

The application of microfluidic technologies to microalgal research is particularly appealing since these approaches allow the precise control of the extracellular environment and offer a high-throughput approach to studying dynamic cellular processes. To expand the portfolio of applications, here we present a droplet-based microfluidic method for analysis and screening of Phaeodactylum tricornutum and Nannochloropsis gaditana, which can be integrated into a genetic transformation workflow. Following encapsulation of single cells in picolitre-sized droplets, fluorescence signals arising from each cell can be used to assess its phenotypic state. In this work, the chlorophyll fluorescence intensity of each cell was quantified and used to identify populations of P. tricornutum cells grown in different light conditions. Further, individual P. tricornutum or N. gaditana cells engineered to express green fluorescent protein were distinguished and sorted from wild-type cells. This has been exploited as a rapid screen for transformed cells within a population, bypassing a major bottleneck in algal transformation workflows and offering an alternative strategy for the identification of genetically modified strains.

Description

Keywords

Fluorescence detection, Microalgae, Microdroplets, Microfluidics, Screening for transformants

Journal Title

Algal Res

Conference Name

Journal ISSN

2211-9264
2211-9264

Volume Title

56

Publisher

Elsevier BV

Rights

All rights reserved
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/I00680X/1)
Biotechnology and Biological Sciences Research Council (BB/L014130/1)
BBSRC (via University College London (UCL)) (FCAGF)
Biotechnology and Biological Sciences Research Council (BB/M50287X/1)
Biotechnology and Biological Sciences Research Council (BB/R021694/1)
Biotechnology and Biological Sciences Research Council (BB/L002957/1)
BBSRC (1838346)