Cryogenic contrast-enhanced microCT enables nondestructive 3D quantitative histopathology of soft biological tissues


Being able to visualize the intricate 3D structure of biological tissues at the micro-scale (i.e., microstructure) is crucial to fully understand their functioning, as well as the impact of pathologies and their treatments.

However, conventional techniques such as classical 2D histological sectioning only partially reveals this complex 3D microstructure of tissues.

In this regard, X-ray based 3D histology, using contrast-enhanced microCT (CECT), can provide important complementary microstructural information.

In our publication in Nature Communications (Nature Portfolio), we present an extension to conventional CECT, termed Cryogenic Contrast-Enhanced MicroCT (cryo-CECT), which combines sample staining with subsequent freezing at the optimal freezing rate, followed by microCT imaging in the frozen state. Using cryo-CECT we were able to substantially improve the visualization of soft biological tissue constituents, such as individual muscle & collagen fibers. To demonstrate the histopathological potential of cryo-CECT, we applied our technique to a disease heart model (TAC). Thanks to cryo-CECT, we were able to analyze, in an unprecedented manner, the orientation and diameter of individual muscle fibers in the entire heart, as well as the 3D localization of fibrotic regions. We foresee further applications of cryo-CECT in the optimization of tissue/food preservation and donor banking, showing that cryo-CECT also has clinical and industrial potential.

Published on October 27, 2022