Abstract
Optical mapping is a high-resolution fluorescence imaging technique, which provides highly detailed visualizations of the electrophysiological wave phenomena, which trigger the beating of the heart. Recent advancements in optical mapping have demonstrated that the technique can now be performed with moving and contracting hearts and that motion and motion artifacts, once a major limitation, can now be overcome by numerically tracking and stabilizing the heart's motion. As a result, the optical measurement of electrical activity can be obtained from the moving heart surface in a co-moving frame of reference and motion artifacts can be reduced substantially. The aim of this study is to assess and validate the performance of a 2D marker-free motion tracking algorithm, which tracks motion and non-rigid deformations in video images. Because the tracking algorithm does not require markers to be attached to the tissue, it is necessary to verify that it accurately tracks the displacements of the cardiac tissue surface, which not only contracts and deforms, but also fluoresces and exhibits spatio-temporal physiology-related intensity changes. We used computer simulations to generate synthetic optical mapping videos, which show the contracting and fluorescing ventricular heart surface. The synthetic data reproduces experimental data as closely as possible and shows electrical waves propagating across the deforming tissue surface, as seen during voltage-sensitive imaging. We then tested the motion tracking and motion-stabilization algorithm on the synthetic as well as on experimental data. The motion tracking and motion-stabilization algorithm decreases motion artifacts approximately by 80% and achieves sub-pixel precision when tracking motion of 1–10 pixels (in a video image with 100 by 100 pixels), effectively inhibiting motion such that little residual motion remains after tracking and motion-stabilization. To demonstrate the performance of the algorithm, we present optical maps with a substantial reduction in motion artifacts showing action potential waves propagating across the moving and strongly deforming ventricular heart surface. The tracking algorithm reliably tracks motion if the tissue surface is illuminated homogeneously and shows sufficient contrast or texture which can be tracked or if the contrast is artificially or numerically enhanced. In this study, we also show how a reduction in dissociation-related motion artifacts can be quantified and linked to tracking precision. Our results can be used to advance optical mapping techniques, enabling them to image contracting hearts, with the ultimate goal of studying the mutual coupling of electrical and mechanical phenomena in healthy and diseased hearts.
Highlights
Optical mapping is a high-resolution fluorescence imaging technique, which is widely used in basic cardiovascular science (Herron et al, 2012)
Due to the shifts of the tissue and a deallocation or dissociation of tissue regions and their corresponding pixel on the camera sensor measuring the tissue region during this pixel-based measurement, the video contains dissociation-related motion artifacts. Such dissociation motion artifacts appear as a network-like, tile-shaped spatial pattern with black-and-white deflections superimposing and distorting the action potential wave pattern, which can be seen in the upper image sequence
We further found that the motion tracking algorithm is robust against fluorescence intensity fluctuations which are caused by electrical activity
Summary
Optical mapping is a high-resolution fluorescence imaging technique, which is widely used in basic cardiovascular science (Herron et al, 2012). It employs optical probes or fluorophores, excitation light, high-speed cameras and filtering equipment and is typically used to image the electrophysiological activity that triggers the beating of the heart. With voltage- and calciumsensitive dyes, for instance, it is possible to image action potential and calcium waves propagating across the heart surface in great detail and at very high speeds. Recent developments have demonstrated that action potential and calcium waves can be imaged as they propagate across the strongly contracting and deforming heart surface (Zhang et al, 2016; Christoph et al, 2017, 2018). The simultaneous imaging of both the heart’s contractile motion and the electrochemical processes that generate the heart’s contractions is pivotal for a better understanding of the heart’s electrophysiology and mechanics and their mutual coupling
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