The organization of Pinus sylvestris pollen tubes during growth was studied by video microscopy of living cells and by electron microscopy after freeze-fixation and freeze-substitution (FF-FS). Pollen germinated and the tubes grew slowly for a total period of about 7 days. Some of the grains formed two tubes, while 10-50% of the tubes ramified. These features are in accordance with development in vivo. The cytoplasmic hyaline cap at the tip disappeared during the 2nd or 3rd day of culture. Ag gregates of starch grains progressively migrated from the grain into the tube and later into the branches. Vacuoles first appeared at day 2 and eventually filled large parts of the tube. The tube nucleus was located at variable dis tances from the tip. Some of the organelles showed lin ear movements in a mostly circulatory pattern, but the majority of the organelles showed brownian-like move ments. Rhodamine-phalloidin-stained actin filaments had a gross axial orientation and were found throughout the tube including at the tip. The ultrastructure of pollen tubes was well preserved after FF-FS, but signs of shrinkage were visible. The secretory vesicles in grow ing tips were not organized in a vesicle cone, and coated pits had a low density with only local accumulations, which is in accordance with slow growth. The mitochon dria contained small cristae and a darkly stained matrix and were located more towards the periphery of the tube, indicating low respiratory activity and low oxygen lev els. The dictyosomes carried typical trans-Golgi net works, but some contained less than the normal number o f cisternae. Other elements of the cytoplasm were irreg ularly spaced rough endoplasmic reticulum, many multivesicular bodies, lipid droplets and two types of vacu oles. The typical organization associated with tip growth in angiosperm pollen tubes, e.g. Ni.coti