3-D image analysis system and megakaryocyte quantitation.

Abstract

A quasi-automatic computer image analysis system has been developed for 3-D reconstruction of stained serial sections and implemented on an IBAS system. Some new automatic image analysis techniques have been designed and incorporated into the system. For image segmentation, a transition region determination based thresholding method is introduced. Neither histogram calculation nor empirical parameters are needed in the automatic threshold selection. A two step 3-D reconstruction procedure--symbolic and pictorial reconstructions--is designed to improve the flexibility and the computational capability of the system. The global level registration and local level registration are separated. The former consists of establishing the relationship among a large numbers of profile pairs dispersed in adjacent sections. A pattern matching method based on pattern recognition principles is devised to exploit the information about the statistical character of mismatch caused by deformation of sections and about the relationship of nearby objects. For the latter, an equivalent elliptical approximation method based on the physical theory of the rotation of rigid bodies is proposed. The system has been used for 3-D reconstruction and quantitation of megakaryocytes in human bone marrow tissue. Features about individual 3-D megakaryocyte cell and the spatial distribution of megakaryocytes are determined. The latter is a new contribution to megakaryocyte quantitation and is not possible by using conventional stereologic techniques. These experimental results have demonstrated the ability of the system to perform quantitative analysis.