Positive Dielectrophoresis-based Raman-activated Droplet Sorting for Culture-free and Label-free Screening of Enzyme Function in Vivo

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2020 Aug 22

PMID 32821836

Citations 24

Authors

Xixian Wang

Yi Xin

Lihui Ren

Zheng Sun

Pengfei Zhu

Yuetong Ji

Chunyu Li

Jian Xu

Bo Ma

Affiliations

Soon will be listed here.

Abstract

The potential of Raman-activated cell sorting (RACS) is inherently limited by conflicting demands for signal quality and sorting throughput. Here, we present positive dielectrophoresis-based Raman-activated droplet sorting (pDEP-RADS), where a periodical pDEP force was exerted to trap fast-moving cells, followed by simultaneous microdroplet encapsulation and sorting. Screening of yeasts for triacylglycerol (TAG) content demonstrated near-theoretical-limit accuracy, ~120 cells min throughput and full-vitality preservation, while sorting fatty acid degree of unsaturation (FA-DU) featured ~82% accuracy at ~40 cells min. From a yeast library expressing algal diacylglycerol acyltransferases (DGATs), a pDEP-RADS run revealed all reported TAG-synthetic variants and distinguished FA-DUs of enzyme products. Furthermore, two previously unknown DGATs producing low levels of monounsaturated fatty acid-rich TAG were discovered. This first demonstration of RACS for enzyme discovery represents hundred-fold saving in time consumables and labor versus culture-based approaches. The ability to automatically flow-sort resonance Raman-independent phenotypes greatly expands RACS' application.

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