Coordinated three-dimensional reconstruction from serial sections at macroscopic and microscopic levels of...

This report describes procedures we have developed for obtaining correlated quantitative structural information at two very different levels of resolution. Accurate reconstructions of entire organs and samples of tissue within organs were produced in correct scalar and topographical relationship using computer-assisted techniques. A specially designed sectioning apparatus, a macrovibratome, was used to section serially the ventricles of the human heart macroscopically. Photographs were taken of every slice. A tissue block excised from a slice at a specified locus in the left ventricular wall was embedded in plastic; serial-3 micron sections were cut in each of two orthogonal orientations. Photomicrographs were taken by semi-automated microscopy. Images of both macroscopic and microscopic sections were projected onto a bitpad and manually digitized. The resulting tables of x-, y-, and z-coordinates were reassembled on a VAX 11/750 computer, then transferred to a high-performance graphics workstation and displayed as three-dimensional images. Microscopic images were shown in the correct reference frame with respect to the macroscopic (parent) structure.