Fluorescence Activated Cell Sorting of Shoot Apical Meristem Cell Types

Growing tips of plants harbor a set of stem cells in structures called shoot apical meristems (SAMs) which provide cells for development of aboveground biomass. Despite a periodic differentiation of stem cell progenitors into leaves, the stem cell pool remains constant over time. Genetic analysis has revealed molecular pathways involved in stem-cell specification, cell division patterns, and organ differentiation. Stem cells within SAMs are few in number, which imposes a limitation to the experimental approaches that can be used for deciphering the gene regulatory networks that underlie cell fate transitions. Here, I provide detailed experimental protocols for the protoplasting and subsequent purification through cell sorting of SAM cells, which allows genome-wide analyses of gene expression patterns at a single cell-type resolution.