TY - CHAP
T1 - Flow Cytometry and Sorting in Arabidopsis
AU - Galbraith, David W.
AU - Sun, Guiling
N1 - Funding Information:
Part of the development of the methods described in this chapter involved support to DG from the NSF Plant Genome Research program, and the National Institutes of Health. DG also acknowledges on-going support from the USDA through the University of Arizona College of Agriculture and Life Sciences. We also acknowledge support from the Chinese National Key Research and Development Program (2OI6YFDOl0l006), the National Natural Science Foundation of China (31770300), the Program for Innovative Research Teams in Science and Technology at a University of Henan Province (18IRTSTHNO23), and the 111 Project (D16014) of China. We finally thank Lisa Villalobos-Menuey (Beckman-Coulter), Laurie Appling (SONY), and Matt Alexander and Alex Rodriguez (BioRad), for valuable assistance in generating the data of Figs. 2, 3, and 4.
Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2021
Y1 - 2021
N2 - Flow cytometry and sorting represents a valuable and mature experimental platform for the analysis of cellular populations. Applications involving higher plants started to emerge around 40 years ago and are now widely employed both to provide unique information regarding basic and applied questions in the biosciences and to advance agricultural productivity in practical ways. Further development of this platform is being actively pursued, and this promises additional progress in our understanding of the interactions of cells within complex tissues and organs. Higher plants offer unique challenges in terms of flow cytometric analysis, first since their organs and tissues are, almost without exception, three-dimensional assemblies of different cell types held together by tough cell walls, and, second, because individual plant cells are generally larger than those of mammals. This chapter, which updates work last reviewed in 2014 [Galbraith DW (2014) Flow cytometry and sorting in Arabidopsis. In: Sanchez Serrano JJ, Salinas J (eds) Arabidopsis Protocols, 3rd ed. Methods in molecular biology, vol 1062. Humana Press, Totowa, pp 509–537], describes the application of techniques of flow cytometry and sorting to the model plant species Arabidopsis thaliana, in particular emphasizing (a) fluorescence labeling in vivo of specific cell types and of subcellular components, (b) analysis using both conventional cytometers and spectral analyzers, (c) fluorescence-activated sorting of protoplasts and nuclei, and (d) transcriptome analyses using sorted protoplasts and nuclei, focusing on population analyses at the level of single protoplasts and nuclei. Since this is an update, details of new experimental methods are emphasized.
AB - Flow cytometry and sorting represents a valuable and mature experimental platform for the analysis of cellular populations. Applications involving higher plants started to emerge around 40 years ago and are now widely employed both to provide unique information regarding basic and applied questions in the biosciences and to advance agricultural productivity in practical ways. Further development of this platform is being actively pursued, and this promises additional progress in our understanding of the interactions of cells within complex tissues and organs. Higher plants offer unique challenges in terms of flow cytometric analysis, first since their organs and tissues are, almost without exception, three-dimensional assemblies of different cell types held together by tough cell walls, and, second, because individual plant cells are generally larger than those of mammals. This chapter, which updates work last reviewed in 2014 [Galbraith DW (2014) Flow cytometry and sorting in Arabidopsis. In: Sanchez Serrano JJ, Salinas J (eds) Arabidopsis Protocols, 3rd ed. Methods in molecular biology, vol 1062. Humana Press, Totowa, pp 509–537], describes the application of techniques of flow cytometry and sorting to the model plant species Arabidopsis thaliana, in particular emphasizing (a) fluorescence labeling in vivo of specific cell types and of subcellular components, (b) analysis using both conventional cytometers and spectral analyzers, (c) fluorescence-activated sorting of protoplasts and nuclei, and (d) transcriptome analyses using sorted protoplasts and nuclei, focusing on population analyses at the level of single protoplasts and nuclei. Since this is an update, details of new experimental methods are emphasized.
KW - Agnostic sampling
KW - Flow cytometry
KW - Fluorescent proteins
KW - Gene expression
KW - Next-generation sequencing
KW - Nucleus
KW - Protoplasts
KW - Sorting
KW - Spectral analysis
KW - Transcription
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U2 - 10.1007/978-1-0716-0880-7_12
DO - 10.1007/978-1-0716-0880-7_12
M3 - Chapter
C2 - 33175382
AN - SCOPUS:85096060414
T3 - Methods in Molecular Biology
SP - 255
EP - 294
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -