Automated 3-D montage synthesis from laser-scanning confocal images: application to quantitative tissue-level cytological analysis.

This paper presents a landmark based method for efficient, robust, and automated computational synthesis of high-resolution, two-dimensional (2-D) or three-dimensional (3-D) wide-area images of a specimen from a series of overlapping partial views. The synthesized image is the set union of the areas or volumes covered by the partial views, and is called the "montage." This technique is used not only to produce gray-level montages, but also to montage the results of automated image analysis, such as 3-D cell segmentation and counting, so as to generate large representations that are equivalent to processing the large wide-area image at high resolution. The method is based on computing a concise set of feature-tagged landmarks in each partial view, and establishing correspondences between the landmarks using a combinatorial point matching algorithm. This algorithm yields a spatial transformation linking the partial views that can be used to create the montage. Such processing can be a first step towards high-resolution large-scale quantitative tissue studies. A detailed example using 3-D laser-scanning confocal microscope images of acriflavine-stained hippocampal sections of rat brain is presented to illustrate the method.