Graham Rykiel (1), Claudia S. López (1,2), Jessica L. Riesterer (1,2), Ian Fries (1), Sanika Deosthali (1), Katherine Courchaine (1), Alina Maloyan (3), Kent Thornburg (3), Sandra Rugonyi (1,3)
eLife, 9, October 2020. DOI: 10.7554/eLife.58138

Abstract

Cardiac pumping depends on the morphological structure of the heart, but also its subcellular (ultrastructural) architecture, which enables cardiac contraction. In cases of congenital heart defects, localized ultrastructural disruptions 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, to assess malformations; and its ultrastructure, to assess organelle 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). In a small animal model (chicken embryo), we achieved uniform fixation and staining of the whole heart, without losing ultrastructural preservation on the same sample, enabling correlative multiscale imaging. Our approach enables multiscale studies in models of congenital heart disease and beyond.

How Our Software Was Used

Dragonfly was used to segment and quantify SBF-SEM images, for automatic and manual clean up, and much more.

Author Affiliation

(1) Biomedical Engineering, Oregon Health & Science University, United States.
(2) Multiscale Microscopy Core, Oregon Health & Science University, United States.
(3) Center for Developmental Health, Knight Cardiovascular Institute, Oregon Health & Science University, United States.