Flow Cytometry and Sorting in Arabidopsis

Flow cytometryFlow cytometry and sortingSorting represents a valuable and mature experimental platform for the analysisSorting of cellular populations. Applications involving higher plantsPlants 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 plantsPlants 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 wallsCell walls, and, second, because individual plant Plants 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 cytometryFlow cytometry and sortingSorting to the model plant Plants 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 sortingSorting of protoplastsProtoplasts and nuclei, and (d) transcriptome analyses using sorted protoplastsProtoplasts and nuclei, focusing on population analyses at the level of single protoplastsProtoplasts and nuclei. Since this is an update, details of new experimental methods are emphasized.