Fig. 5. Nile Red staining (orange colour) of lipid and membranes at various times during the regeneration of sub-protoplast. (A) Active movement of lipid material towards the interior of the protoplasmic mass occurred during the first few minutes. (B) The swirling movement of the cell organelles stopped and lipid material was localized to the centre of the protoplasmic mass. (C) The sub-protoplast became spherical and covered with primary envelope, but the envelope is not stained at this stage. (D) The lipid material begins to be incorporated into the envelope at 1 hour after wounding. (E-F) Strong labelling of the primary envelope and secondary membrane. Bars, 10 µm.