In plants, the morphological diversity of leaves is largely determined by cell division, especially cell division orientation. Whereas cell division itself is easily monitored, the detection and quantification of cell division orientation are difficult. The few existing methods for detection and quantification of cell division orientation are either inefficient or laborious. Here, we describe a pulse-chase strategy using a 5-ethynyl-2’-deoxyuridine (EdU) labeling assay. Plant tissues are first incubated with EdU for a short period (pulse), followed by a long incubation without EdU (chase). Using this method, the positions of daughter cells are easily detected and can be used to quantify cell division orientation. Our protocol is rapid and very efficient for quantitative analysis of cell division orientation, and can be applied to both model and non-model plant species.
Graphic abstract Plant cell division pairs clearly visualized by a pulse-chase EdU method