4D Imaging of heart vaso-architecture after myocardial infarction.

Cardiovascular diseases remain the number one cause of death globally. There is an ongoing desire to study the distribution and structural changes of the vaso-architecture in the diseased heart in cardiovascular research groups all over the world.

The ability to acquire high resolution 3D-images of the heart vasculature enables to study heart diseases more in detail and eventually obtain interesting new findings and new treatments. In this work, we introduce a pipeline for high resolution 3D-imaging of the changes in mouse heart vasculature after a myocardial infarction is produced with Single Plane Illumination Microscopy (SPIM).

To achieve high resolution 3D-images, protocols for optical tissue clearing (CUBIC tissue clearing technique) were combined with vasculature labelling methods (IHC and intravenous perfused lectin), enabling the visualization for the very first time of the whole heart vasculature.

We here also describe the methods used for image pre-processing of the acquired data, mainly for correction of SPIM-image artifacts and for segmentation of the structures of interest.

Finally, the analysis of the changes in vasculature between healthy hearts with the different stages of chronic myocardial infarction (7, 14 and 28 days post-infarction) will provide us a tool to know how this disease affects not only to infarcted region but to the whole heart volume.