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In vivo AFM imaging directly in anesthetized living rats. (a) Schematic showing the custom surgical platform that matches a rat body. The luminal side of blood vessel is surgically exposed upwards and is subject to AFM imaging in the center of the hollow imaging dish. (b) Schematic anatomy of the blood vessels. It contains three coats: (i) the inner coat (tunica intima) consists of a single layer of endothelial cells applied to the internal elastic lamina; (ii) the middle coat (tunica media) contains primarily smooth muscle cells and elastic tissue fibers; and (iii) the outer coat (tunica adventitia) embeds the vessel in its surroundings and consists mainly of connective tissue. (c) and (d) show in vivo AFM topographic images of aortic intima, and spontaneous fenestration of aortic intima, respectively. (e) The zoomed-in AFM image of the fenestration in (d), showing fibrous network structure inside the fenestration, mostly belonging to internal elastic lamina. Scale bar, in (c) and (d) and in (e).
Visualization of nanomechanical properties of aortic intima. [(a)–(d)] Nanomechanical characterization of normal endothelia cells on the surface of aortic intima. [(e)–(h)] Nanomechanical characterization of a fenestration on the surface of aortic intima. Topography images [(a) and (e)], elasticity maps [(b) and (f)], energy dissipation maps [(c) and (g)], and representative force-indentation curves [(d) and (h)] in the same row are obtained simultaneously with force-volume imaging. The corresponding spatial positions of the representative force-indentation curves are marked by star and labeled by numbers in (a) and (e), respectively. The elasticity maps are shown as Young’s modulus calculated from linear fitting of Hertz model to force curves at low indentation (50 nm). The energy dissipation maps show the total works performed during the force increasing from 0 to 5 nN. .
In vivo changes of elasticity of blood vessels in response to drug stimulations. The real-time variation of Young’s modulus of endothelial cells in vivo under NG-induced vasodilation (from ) followed by NE-induced vasoconstriction (from ). Modulus as a function of time is measured on the same topography position with translational error of 100 nm.
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