Cell-accurate optical mapping across the entire developing heart

Abstract

Organogenesis depends on orchestrated interactions between individual cells and morphogenetically relevant cues at the tissue level. This is true for the heart, whose function critically relies on well-ordered communication between neighboring cells, which is established and fine-tuned during embryonic development. For an integrated understanding of the development of structure and function, we need to move from isolated snap-shot observations of either microscopic or macroscopic parameters to simultaneous and, ideally continuous, cell-to-organ scale imaging. We introduce cell-accurate three-dimensional Ca2+-mapping of all cells in the entire electro-mechanically uncoupled heart during the looping stage of live embryonic zebrafish, using high-speed light sheet microscopy and tailored image processing and analysis. We show how myocardial region-specific heterogeneity in cell function emerges during early development and how structural patterning goes hand-in-hand with functional maturation of the entire heart. Our method opens the way to systematic, scale-bridging, in vivo studies of vertebrate organogenesis by cell-accurate structure-function mapping across entire organs.

Single cell determination of cardiac microtissue structure and function using light sheet microscopy

“…While single planes within tissues can be imaged at 20Hz, it takes several seconds to image an entire z-stack at the fastest acquisition rate. This acquisition speed limitation could potentially be overcome by acquiring high-speed single plane time series every few micrometers apart within the tissue followed by post-imaging stitching of these frames, ultimately constructing a 3D network of functional propagation-a method known as post acquisition synchronization 41 . Another challenge is that additional indepth quantitative assessments of calcium handling properties could not be determined using the Zeiss z.1 microscope.…”

Hybrid optical gating for long-term 3D time-lapse imaging of the beating embryonic zebrafish heart

“…However, to date, only some aspects of this problem have been solved at any one time. Imaging of the developing fish and chick heart has been previously reported for: single 3D snapshots at selected intervals [11], 3D images of a single cardiac cycle [12,13], sampling of a limited number of time points during development [9,14], and 2D time-lapse video [15]. Temporarily stopping the heart using high dose anaesthetic has also been used to acquire 3D image sets of the heart [16] but such approaches may significantly alter the physiological a c b Alternative heart green?…”

Cyborg Organoids: Implantation of Nanoelectronics via Organogenesis for Tissue-Wide Electrophysiology

“…heart and brain) rely on spatiotemporally orchestrated communication of heterogenous cells to generate whole-organ functions. 1,2 Thus, understanding development, mechanism and diseases of these organs requires system-level mapping of the cellular activities with high spatiotemporal resolutions across their entire 3D volumes over a substantial time window. [3][4][5] Recently, tremendous progresses have miniaturized state-of-the-art electrophysiological tools into micro/nanoscales 6,7 with soft and multiplexed electronic units, 8,9 which have significantly reduced tissue disruptions while maintaining the unmatched spatiotemporal resolution for recording and manipulating.…”