Deformation‐corrected computer‐aided three‐dimensional reconstruction of immunohistochemically stained organs: Application to the rat heart during early organogenesis

Abstract

The application of a computer-assisted, three-dimensional reconstruction procedure for serial sections to embryonic rat hearts during the period of cardiac looping and compartmentalization is described. The procedure relies on immunohistochemical staining for the introduction of selective contrast and on episcopic and diascopic images of each of the sections for alignment and correction of compression due to sectioning. Episcopic (reference) images are taken from the embedding block just before the cutting of a slice and are still aligned and undeformed. Diascopic images are taken from the sections after immunohistochemical processing and, hence, contain selective contrast but are deformed and no longer aligned. The three-dimensional images are visualized as shaded voxel models. This approach allowed the unequivocal delineation of the developing myocardium and the inspection of its changing architecture both from the outside and from within. Furthermore, it allowed a quantification of myocardial volume. Because standardized and hence comparable views of three different stages were generated, changes in the shape of the cardiac loop, the atria, and the ventricles as well as changes in the position of the atrioventricular canal and interventricular foramen could be accurately described. Characteristic changes in the position of both the right ventricle and the atrioventricular canal that are essential for the formation of a correctly functioning four-chambered heart could be observed. These changes in shape occur while the myocardial size increases dramatically.

Evaluation of two 3D virtual computer reconstructions for comparison of cleft lip and palate to normal fetal microanatomy

“…The absolute orientation of each frame is established by a stochastic analysis based on anatomical landmarks. Episcopic pictures taken from the microtome block and alignment algorithm can be used alternatively (Laan et al, 1989;Lozanoff, 1992).…”

“…No matter how much error is tolerated, the reconstruction must yield a result adequately exact and detailed to comply with the objectives of faithfully representing the spatial and volume relationships for a realistic mesh generation and finite-element analysis (FEA), deformation calculation, and education (i.e., optimum comprehensibility). These issues have been reviewed elsewhere (Prothero and Prothero, 1986;Brä ndle, 1989;Deverell et al, 1989Deverell et al, , 1993Hara et al, 1989;Laan et al, 1989;Lozanoff and Deptuch 1991;McLean and Prothero, 1991;Ongaro et al, 1991;Lozanoff, 1992;Marco and Leith, 1992;Moss, 1992;Vuillemin et al, 1992;Clarke et al, 1993;Hibbard et al, 1993). The use of computers in 3D reconstruction began more than 20 years ago (Gaunt and Gaunt, 1978;Huijsmans et al, 1986) and fundamental algorithms have been outlined (Johnson and Capowski, 1985;Moss, 1992;Kvasnicka and Thiele, 1995).…”

Design and implementation of a database and program for 3D reconstruction from serial sections: A data‐driven approach

“…It may be clear from the description of the program that a simple solution has been chosen for the alignment procedure (see above, "Reconstruction and Realignment of the Data"). As more elaborate methods become available, (Laan et al, 1989;Merikel, 1988;Verbeek et al, 1990bVerbeek et al, , 1991aVerbeek, 1992), these can be embedded easily in the database structure and into the reconstruction program.…”

“…For a lot of reconstructions, realignment based on rigid transformations is sufficient. When more reference marks are available, more sophisticated realignment methods (Laan et al, 1989;Merikel, 1988;Verbeek et al, 1990bVerbeek et al, , 1991aVerbeek, 1992) may be used. These methods are currently not available to this program.…”

Three-dimensional reconstruction of histological sections using modern product-design software

“…No matter how much error is tolerated, the reconstruction must yield a result adequate to answer the scientific question posed. These issues have been reviewed elsewhere; see, e.g., Burston and Thurley (1957); Prothero and Prothero (1986); Brä ndle (1989); Deverell et al (1989;; Hara et al (1989); Laan (1989); Lozanoff and Deptuch (1991); McLean and Prothero (1991); Ongaro et al (1991); Lozanoff (1992); Marco and Leith (1992); Moss (1992); Vuillemin et al (1992); Clarke et al (1993); Hibbard et al (1993).…”