Publications
Resources / Publications
Graham Rykiel (1), Claudia S. López (2), Jessica L. Riesterer (2), Ian Fries, Sanika Deosthali, Katherine Courchaine (1), Alina Maloyan (1), Kent Thornburg (1), Sandra Rugonyi (1)
Cold Spring Harbor Laboratory Press, April 2020. DOI: 10.1101/2020.04.22.055418
Developmental Biology
Efficient cardiac pumping depends on the morphological structure of the heart, but also on its sub-cellular (ultrastructural) architecture, which enables cardiac contraction. In cases of congenital heart defects, localized sub-cellular disruptions in architecture that increase the risk of heart failure are only starting to be discovered. This is in part due to a lack of technologies that can image the three dimensional (3D) heart structure, assessing malformations; and its ultrastructure, assessing disruptions. We present here a multiscale, correlative imaging procedure that achieves high-resolution images of the whole heart, using 3D micro-computed tomography (micro-CT); and its ultrastructure, using 3D scanning electron microscopy (SEM). This combination of technologies has not been possible before in imaging the same cardiac sample due to the heart large size, even when studying small fetal and neonatal animal models (~5×5×5mm3). Here, we achieved uniform fixation and staining of the whole heart, without losing ultrastructural preservation (at the nm resolution range). Our approach enables multiscale studies of cardiac architecture in models of congenital heart disease and beyond.
Dragonfly was used to segment (delineate) from SBF-SEM images the cell nuclei, myofibrils, mitochondria, and the extracellular space, to better visualize and quantify the cardiac ultrastructure.
(1) Center for Developmental Health, Knight Cardiovascular Institute.
(2) Multiscale Microscopy Core, Oregon Health & Science University, Portland Oregon, USA.
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