A method for 3D reconstruction of neuronal processes using semithin serial sections displayed as a cinematographic sequence.

In order to study the organization and distribution of dendrites and axons in the cerebral cortex, we have developed a computer-assisted method for 3D reconstruction of neuronal processes based on serial light microscopic images displayed as a continuous sequence. A series of tangential sections (0.65 micron thick) through rat parietal cortex was aligned, digitized into the computer and then used to build a sequence (stack) of images which was stored to a digital real-time video disk. Apical dendrites located in dendritic bundles in laminae III and IV were traced through the sequence. Two tracing modes were tested: (1) cinematographic mode, in which the image stack was displayed continuously and automatically by the computer at various preset speeds (max. speed: 25 images/s) and (2) stepping mode, in which the interval between each image was varied manually according to the choice of the operator. Coordinates were stored in a database and used to build a 3D reconstruction where apical dendrites were displayed as wires or tubes. Tracing in cinematographic mode was about 3 times faster than tracing in stepping mode. We believe that the former mode exploits the built in 'filtering' capacity of the visual system to perform temporal averaging.