Separation and pseudo-coloring of high-speed bright-field microscopy images of the beating embryonic heart

High-speed, bright-field (BF) microscopy of the beating embryonic zebrafish heart reveals both static background structures as well as the rapid motion of cardiac tissues and red blood cells (RBCs). However, all structures contribute to BF image contrast in a similar way, making labeling and subsequent analysis of these images difficult. Here, we report on our progress to separate cardiac BF images into three separate channels, based on the periodic, aperiodic, or static motion patterns respective to each of the cardiac structures. Specifically, we improve on a previously developed algorithm that was limited to two-class separation only. We show that the three extracted channels can be recombined into a pseudo-colored sequence in which RBCs as well as heart wall and background structures appear in different colors. We anticipate that these improvements will facilitate quantitative characterization of heart function during normal and abnormal cardiac development.