3D reconstruction of chloroplasts and their ultrastructure using ultra-thin-serial-sections

Reconstruction of fine structures of plant organelles can not be done applying light microscopy. The resolution of this technique is too poor, therefore Transmission Electron Microscopy (TEM) has to be used. However, it is impossible to evaluate larger parts of the cells or whole cell organelles because specimens are too thin for these topics. This is also the reason why computer-tomography is impossible. Therefore series of ultra-thin-sections were used. Black and white images of TEM-preparations were transformed into pixel-images with a scanner for further analysis with a computer. After this procedure the images were transformed into vector-graphs. Now it was possible to reconstruct organelles in a 3D view and to calculate areas and volumes. Illuminated spinach leaves were used as objects for evaluation. The results of the 3D reconstruction of spinach chloroplasts with their ultrastructures and the calculations of the volumes of their components are shown in this paper.