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. 2022 Feb 8;7(3):e153913. doi: 10.1172/jci.insight.153913

Figure 1. Optical mapping and tissue clearing pipeline to align electrical and structural maps.

Figure 1

(A) Schematic of optical mapping, clearing, imaging, and automated feature tracing steps in the alignment pipeline. (B and C) Bright-field image taken simultaneously with optical action potential map showing activation in sinus rhythm. (D) Maximum intensity projection (MIP) image of tyrosine hydroxylase–positive (TH-positive) nerve fibers on the ventral surface of the same heart after IHC, tissue clearing, and confocal imaging. (E and F) High-magnification images of the boxed region in D, with TH staining alongside nerve-fiber tracing by computer vision, color-coded by fiber diameter. (G and H) Venous bifurcations (magenta points) on MIP confocal shell image of a cleared heart alongside bright-field image of same heart were used as fiducial anchors for alignment. (I) Automated global nerve-fiber tracing aligned with bright-field image allows spatial correlation with optical action potential data. Scale bars: 1 mm (B–D and G–I); 100 μm (E and F).