Title: A Gelatin Microdroplet Platform for High‐Throughput Sorting of Hyperproducing Single‐Cell‐Derived Microalgal Clones

Abstract

Abstract Microalgae are an attractive feedstock organism for sustainable production of biofuels, chemicals, and biomaterials, but the ability to rationally engineer microalgae to enhance production has been limited. To enable the evolution‐based selection of new hyperproducing variants of microalgae, a method is developed that combines phase‐transitioning monodisperse gelatin hydrogel droplets with commercial flow cytometric instruments for high‐throughput screening and selection of clonal populations of cells with desirable properties, such as high lipid productivity per time traced over multiple cell cycles. It is found that gelatin microgels enable i) the growth and metabolite (e.g., chlorophyll and lipids) production of single microalgal cells within the compartments, ii) infusion of fluorescent reporter molecules into the hydrogel matrices following a sol–gel transition, iii) selection of high‐producing clonal populations of cells using flow cytometry, and iv) cell recovery under mild conditions, enabling regrowth after sorting. This user‐friendly method is easily integratable into directed cellular evolution pipelines for strain improvement and can be adopted for other applications that require high‐throughput processing, e.g., cellular secretion phenotypes and intercellular interactions.

Authors:

Li, Ming  ^[1];

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van Zee, Mark ^[2]; Riche, Carson T. ^[2]; Tofig, Bobby ^[3]; Gallaher, Sean D. ^[4]; Merchant, Sabeeha S. ^[4]; Damoiseaux, Robert ^[5]; Goda, Keisuke ^[6]; Di Carlo, Dino ^[7]

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+ Show Author Affiliations

1. Department of Bioengineering University of California, Los Angeles Los Angeles CA 90095 USA, School of Engineering Macquarie University Sydney NSW 2122 Australia
2. Department of Bioengineering University of California, Los Angeles Los Angeles CA 90095 USA
3. California NanoSystems Institute University of California, Los Angeles Los Angeles CA 90095 USA
4. Department of Chemistry and Biochemistry University of California, Los Angeles Los Angeles CA 90095 USA
5. California NanoSystems Institute University of California, Los Angeles Los Angeles CA 90095 USA, Department of Molecular and Medicinal Pharmacology University of California, Los Angeles Los Angeles CA 90095 USA
6. Department of Chemistry University of Tokyo Tokyo 113‐8655 Japan, Japan Science and Technology Agency Kawaguchi 332‐0012 Japan, Department of Electrical Engineering University of California, Los Angeles Los Angeles CA 90095 USA
7. Department of Bioengineering University of California, Los Angeles Los Angeles CA 90095 USA, California NanoSystems Institute University of California, Los Angeles Los Angeles CA 90095 USA, California NanoSystems Institute Jonsson Comprehensive Cancer Centre University of California, Los Angeles Los Angeles CA 90095 USA

Publication Date:

Thu Oct 04 00:00:00 EDT 2018

Sponsoring Org.:

USDOE

OSTI Identifier:

1479571

Grant/Contract Number:  

DE‐FC02‐02ER63421

Resource Type:

Publisher's Accepted Manuscript

Journal Name:

Small

Additional Journal Information:

Journal Name: Small Journal Volume: 14 Journal Issue: 44; Journal ID: ISSN 1613-6810

Publisher:

Wiley Blackwell (John Wiley & Sons)

Country of Publication:

Germany

Language:

English