Fluorescence activated cell sorting—A selective tool for plant cell isolation and analysis

Instrumentation for flow cytometry and sorting is designed around the assumption that samples are single‐cell suspensions. However, with few exceptions, higher plants comprise complex multicellular tissues and organs, in which the individual cells are held together by shared cell walls. Single‐cell...

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Published inCytometry. Part A Vol. 101; no. 9; pp. 725 - 736
Main Authors Antoniadi, Ioanna, Skalický, Vladimír, Sun, Guiling, Ma, Wen, Galbraith, David W., Novák, Ondřej, Ljung, Karin
Format Journal Article
LanguageEnglish
Published Hoboken, USA John Wiley & Sons, Inc 01.09.2022
Wiley Subscription Services, Inc
Subjects
autofluorescence
Best practice
best practices
Cell Biology
Cell suspensions
Cell walls
Cellbiologi
Flow cytometry
Instrumentation
plant flow cytometry and sorting
Protoplasts
viability and integrity
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Summary:Instrumentation for flow cytometry and sorting is designed around the assumption that samples are single‐cell suspensions. However, with few exceptions, higher plants comprise complex multicellular tissues and organs, in which the individual cells are held together by shared cell walls. Single‐cell suspensions can be obtained through digestion of the cells walls and release of the so‐called protoplasts (plants without their cell wall). Here we describe best practices for protoplast preparation, and for analysis through flow cytometry and cell sorting. Finally, the numerous downstream applications involving sorted protoplasts are discussed.
Bibliography:Funding information
Chinese National Key Research and Development Program, the National Natural Science Foundation of China, the Program for Innovative Research Teams in Science and Technology at a University of Henan Province, and the 111 Project of China; Czech Foundation Agency, Grant/Award Number: GA17‐21581Y; Endowment fund of Palacký University in Olomouc; European Molecular Biology Organization, Grant/Award Number: EMBO ASTF 297‐2013; Internal Grant Agency of Palacký University, Grant/Award Number: IGA_PrF_2021_011; Kempestiftelserna; Ministry of Education, Youth and Sport of the Czech Republic via the ERDF project “Plants as a tool for sustainable global development”, Grant/Award Number: CZ.02.1.01/0.0/0.0/16_019/0000827; Plant Fellows (the International Post doc Fellowship Programme in Plant Sciences, Grant/Award Number: 267423; the Knut and Alice Wallenberg Foundation (KAW); the Swedish Governmental Agency for Innovation Systems (Vinnova); the Swedish Research Council; University of Arizona College of Agriculture and Life Sciences
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ISSN:1552-4922
1552-4930
1552-4930
DOI:10.1002/cyto.a.24461