Figure 2
From: A simple way to identify non-viable cells within living plant tissue using confocal microscopy
SYTOX orange staining of non-viable lateral root cap cells. (A) to (D) Enhancer-trap line Q0171 expressing GFP (green) in the columella (c) and lateral root cap (lrc). (A), (C) Overlay projection image of Q0171 stained with propidium iodide. The end of the lateral root cap (box in (A)) is magnified in (C). Propidium iodide stain (red) outlines cells and stains nuclei of non-viable root cap cells (arrows in (C)). (B) CLSM image showing a virtual optical section through the middle of a Q0171 root stained with propidium iodide. To visualise cells in the centre of the root, the root was stained with propidium iodide for an extended time (10 minutes) and therefore cells at the periphery of the root are heavily stained. The lateral root cap is marked with an arrow and a bracket shows the length of the lateral root cap. (D) Overlay projection image of line Q0171 stained with SYTOX orange: non-viable cells are stained selectively. (E) to (H) Overlay projection images of the end of the lateral root cap of wild type roots stained with fluorescein diacetate (labels living cells, shown in cyan) and SYTOX orange (red). (E) Fluorescein diacetate fluorescence. (F) SYTOX orange fluorescence in the same root. (G) Overlay of (E) and (F). (H) Scheme outlining the SYTOX orange stained cells. All roots are 5 days old. Scalebars: 100 μm.