Abstract
The ability to acquire 3D images of the heart and its vasculature at cellular resolution facilitates a more detailed study of many heart diseases. Here, we describe a novel technique to image in 3D the heart vasculature by combining the CUBIC clearing protocol combined with in vivo administration of fluorescent-labeled lectin. The use of these techniques in combination with Selective Plane Illumination Microscopy (SPIM) made it possible to obtain high resolution 3D images of the cardiac vascular tree. This methodological approach may enhance the visualization of 3D images of the cardiac vasculature remodeling associated with coronary disease.
Highlights
Coronary artery disease is accompanied by significant structural changes in the vascular network, including the thickening of large arteries, abnormal lipid deposits on the blood vessel wall and lumen narrowing [1].These pathological changes may lead to a myocardial infarction and eventually to heart failure
We present 3D images of the cardiac vasculature processed with the illumination correction algorithm
CUBIC and BABB are the tissue clearing methods widely preferred over other methods [6, 17]
Summary
Coronary artery disease is accompanied by significant structural changes in the vascular network, including the thickening of large arteries, abnormal lipid deposits on the blood vessel wall and lumen narrowing [1]. These pathological changes may lead to a myocardial infarction and eventually to heart failure. Confocal microscopy is a well-established technique to image and study tissues at cellular and subcellular resolution, and has been used to obtain microvascular imaging of different murine tissues [4]. Overcoming the limited 3D abilities of confocal microscopy, Selective Plane Illumination Microscopy (SPIM) has been utilized to acquire threedimensional information of large intact rodent organs [5]. Protocols to obtain this kind of information in the adult heart are still missing
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