Longitudinal imaging of a 3D model of calcific aortic valve disease, which consisted of co-cultured GFP+ Valve Endothelial Cells (VEC) and Valve Interstitial Cells (VIC), was performed with a combined Optical Coherence Microscopy (OCM), confocal reflectance and fluorescence microscopy system. The acquired confocal volumes depicted the VEC morphological changes and migration as well as collagen fiber alignment. With the aid of computational refocusing and multi-volume processing, the OCM datasets could visualize VIC cell bodies, matrix remodeling, nodule formation and calcific deposits. The complementary information derived using this combined approach could help unravel the cellular mechanisms leading to aortic valve calcification.
A combined 1300 nm Optical Coherence Microscopy (OCM) and 488 nm confocal reflectance/ fluorescence microscopy system was designed to perform high-resolution, high-specificity imaging of collagen-embedded spheroids. Spheroids of Hyaluronic Acid (HA) synthase-overexpressing breast epithelial cells alone, or co-cultured with adipose stromal cells were imaged. The volumes, acquired either after fixing and staining or longitudinally with labeling, enabled the visualization of the spheroid morphology, luminal structures, cellular organization, and collagen matrix remodeling. The morphology and internal lumen structures of spheroids, as large as 500 μm in diameter, could be obtained from the OCM volumes, even in the presence of dense collagen matrix surrounding the spheroids. The confocal stacks provided superior specificity to discriminate cells from the compacted collagen along the spheroid’s periphery, up to a depth of ~120 μm. The combined use of OCM and confocal imaging on these spheroid models has added to our understanding of how HA may contribute to tumor initiation and invasion.
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