Three-dimensional vacuole reconstruction of a vacuolated BobTIP26-1::gfp expressing cell 7 days after subculture. (a) Projection view of 40 confocal serial pictures corresponding to the half depth of the cell (i.e. 40 μm). Bar = 25 μm. (b) 3-D view after isosurface extraction showing the protoplasmic side of the vacuole. (c) Interior view of the vacuole after isosurface extraction. Green and red correspond to the tonoplast and the chloroplasts, respectively. Arrow: nuclear pouch; arrowheads: transvacuolar strands. The missing domains of the tonoplast surface in (b) and (c) result from an under-sampling of confocal images. The rendering of a completely smooth 3-D view would have required use of additional intermediate sections.