Entcheva Emilia, Bien Harold
Department of Biomedical Engineering, Stony Brook University, HSC T18-030, Stony Brook, NY 11794-8181, USA.
Prog Biophys Mol Biol. 2006 Oct;92(2):232-57. doi: 10.1016/j.pbiomolbio.2005.10.003. Epub 2005 Nov 21.
Optical mapping of cardiac excitation using voltage- and calcium-sensitive dyes has allowed a unique view into excitation wave dynamics, and facilitated scientific discovery in the cardiovascular field. At the same time, the structural complexity of the native heart has prompted the design of simplified experimental models of cardiac tissue using cultured cell networks. Such reduced experimental models form a natural bridge between single cells and tissue/organ level experimental systems to validate and advance theoretical concepts of cardiac propagation and arrhythmias. Macroscopic mapping (over >1cm(2) areas) of transmembrane potentials and intracellular calcium in these cultured cardiomyocyte networks is a relatively new development and lags behind whole heart imaging due to technical challenges. In this paper, we review the state-of-the-art technology in the field, examine specific aspects of such measurements and outline a rational system design approach. Particular attention is given to recent developments of sensitive detectors allowing mapping with ultra-high spatiotemporal resolution (>5 megapixels/s). Their interfacing with computer platforms to match the high data throughput, unique for this new generation of detectors, is discussed here. This critical review is intended to guide basic science researchers in assembling optical mapping systems for optimized macroscopic imaging with high resolution in a cultured cell setting. The tools and analysis are not limited to cardiac preparations, but are applicable for dynamic fluorescence imaging in networks of any excitable media.
利用电压敏感染料和钙敏感染料对心脏兴奋进行光学映射,使人们得以独特地洞察兴奋波动力学,并推动了心血管领域的科学发现。与此同时,天然心脏的结构复杂性促使人们利用培养的细胞网络设计简化的心脏组织实验模型。这种简化的实验模型在单细胞与组织/器官水平的实验系统之间架起了一座天然的桥梁,以验证和推进心脏传导及心律失常的理论概念。对这些培养的心肌细胞网络中的跨膜电位和细胞内钙进行宏观映射(覆盖面积超过1平方厘米)是一项相对较新的进展,由于技术挑战,其发展落后于全心脏成像。在本文中,我们回顾了该领域的最新技术,审视了此类测量的具体方面,并概述了一种合理的系统设计方法。我们特别关注了能够实现超高时空分辨率(>5百万像素/秒)映射的灵敏探测器的最新进展。本文还讨论了它们与计算机平台的接口,以匹配新一代探测器特有的高数据通量。这篇批判性综述旨在指导基础科学研究人员组装光学映射系统,以便在培养细胞环境中进行高分辨率的优化宏观成像。这些工具和分析不仅限于心脏标本,还适用于任何可兴奋介质网络中的动态荧光成像。