High resolution scanning and three-dimensional reconstruction of cellular events in large objects during brain development

Abstract

Detailed knowledge of the spatial and temporal interactions of distinct cellular events and of the genes involved in their regulation is a precondition for the understanding of morphogenetic and pathogenetic processes. Here, how patterns of cellular events in large objects can be visualized with the help of the image acquisition system ‘Huge Image’ is demonstrated. Huge images are composed of a multitude of small images scanned with the highest light microscopical resolution. The system is equipped with a programmable autofocus device and permits precise and rapid cytological diagnosis. A vector-based three-dimensional (3-D) reconstruction method which, in future projects, will be combined with ‘Huge Image’, is applied to visualize dynamic interactions between macrophages and the occurrence of apoptotic neuroepithelial cells in the early developing forebrain of Tupaia belangeri (Scandentia). Proportionally correct meshwire surfaces of small and large objects are generated independently of each other. The combined reconstruction of cellular events and large embryonic surfaces can be carried out from only subsets of histological serial sections, and, compared with volume-based systems, with a much lower need for memory. The practicability of our approach is compared with recent other methods used to demonstrate apoptotic patterns.