Monitoring of the process of immersion optical clearing of collagen bundles using optical coherence tomography

Future development of the method of immersion optical clearing of biological tissues–this method is widely used in the study of the morphology and pathologies of tissues in vitro and considered promising for in vivo applications in biophysical research and medicine—requires knowledge of the details of interaction of immersion liquids with the tissue, in particular, the characteristics both of the tissue dehydration process, which is caused by the osmotic effect of the immersion liquid, and the process of diffusion of the immersion agent (IA) into the tissue. The optical properties of skin dermis, eye sclera, tendon, and many other tissues are determined by the properties of collagen bundles, abundant in these tissues. In the present work, a convenient and reliable technique for monitoring the optical properties and geometry of collagen bundles in the course of their immersion clearing in vitro, based on optical coherence tomography (OCT), is proposed. The main advantage of this technique is that it allows one to monitor changes in the geometric and optical properties of the tissue simultaneously, without interrupting the natural course of the immersion clearing process, and to obtain reliable estimates of the characteristic times and rates of both the process of tissue dehydration and process of diffusion of the IA into the tissue.