We propose a methodology for making optimal registration decisions during 3D volume reconstruction in terms of (a) anticipated accuracy of aligned images, (b) uncertainty of obtained results during the registration process, (c) algorithmic repeatability of alignment procedure, and (d) computational requirements. We researched and developed a web-enabled, web services based, data-driven, registration decision support system. The registration decisions include (1) image spatial size (image sub-area or entire image), (2) transformation model (e.g., rigid, affine or elastic), (3) invariant registration feature (intensity, morphology or a sequential combination of the two), (4) automation level (manual, semi-automated, or fully-automated), (5) evaluations of registration results (multiple metrics and methods for establishing ground truth), and (6) assessment of resources (computational resources and human expertise, geographically local or distributed). Our goal is to provide mechanisms for evaluating the tradeoffs of each registration decision in terms of the aforementioned impacts. First, we present a medical registration methodology for making registration decisions that lead to registration results with well-understood accuracy, uncertainty, consistency and computational complexity characteristics. Second, we have built software tools that enable geographically distributed researchers to optimize their data-driven registration decisions by using web services and supercomputing resources. The support developed for registration decisions about 3D volume reconstruction is available to the general community with the access to the NCSA supercomputing resources. We illustrate performance by considering 3D volume reconstruction of blood vessels in histological sections of uveal melanoma from serial fluorescent labeled paraffin sections labeled with antibodies to CD34 and laminin. The specimens are studied by fluorescence confocal laser scanning microscopy (CLSM) images.
We address the problem of automated image alignment for 3D volume reconstruction from stacks of fluorescent confocal laser scanning microscope (CLSM) imagery acquired at multiple confocal depths, from a sequence of consecutive slides. We focus on automated image alignment based on centroid and area shape features by solving feature correspondence problem, also known as Procrustes problem, in highly deformable and ill-conditioned feature space. In result, we compare image alignment accuracy of a fully automated method with registration accuracy achieved by human subjects using a manual alignment method. Our work demonstrates significant benefits of automation for 3D volume reconstruction in terms of accuracy, consistency, and performance time. We also outline the limitations of fully automated and manual 3D volume reconstruction system.
We present a novel semi-automated registration technique for 3D volume reconstruction from fluorescent laser scanning confocal microscope (LSCM) imagery. The developed registration procedure consists of (1) highlighting segmented regions as salient feature candidates, (2) defining two region correspondences by a user, (3) computing a pair of region centroids, as control points for registration, and (4) transforming images according to estimated transformation parameters determined by solving a set of linear equations with input control points. The presented semi-automated method is designed based on our observations that (a) an accurate point selection is much harder for a human than an accurate region (segment) selection, (b) a centroid selection of any region is less accurate by a human than by a computer, and (c) registration based on structural shape of a region rather than on intensity-defined point is more robust to noise and to morphological deformation of features across stacks. We applied the method to image mosaicking and image alignment registration steps and evaluated its performance with 20 human subjects on LSCM images with stained blood vessels. Our experimental evaluation showed significant benefits of automation for 3D volume reconstruction in terms of achieved accuracy, consistency of results and performance time. In addition, the results indicate that the differences between registration accuracy obtained by experts and by novices disappear with an advanced automation while the absolute registration accuracy increases.