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Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes

机译:单个心肌细胞微区钙收缩偶联的多峰二次谐波产生和双光子荧光成像

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The objective of this study was to develop a method for simultaneously measuring the calcium and contraction dynamics of single, live cardiomyocytes at high spatial resolutions. Such measurements are important to investigate local calcium release and the mechanical response at the sarcomere level (i.e. the basic unit of contraction), which have important implications in cardiac dysfunction and arrhythmias in conditions such as hypertension, atrial fibrillation, and myocardial infarction. Here, we describe a multimodal second harmonic generation (SHG) and two photon fluorescence (2PF) microscopy technique that is used to simultaneously measure subsarcomere calcium and contraction events at high spatial and temporal resolutions. The method takes advantage of the label-free nature of SHG for imaging the sarcomeres and the high spatial colocalization of the SHG signal and the fluorescence signal excited from calcium indicators. This microscope was used to measure calcium sparks and waves and associated contractions in subcellular microdomains, leading to the generation of subcellular strain. We anticipate this new imaging tool will play an important role in studying mechanical stress-induced heart disease.
机译:这项研究的目的是开发一种同时以高空间分辨率测量单个活心肌细胞钙和收缩动态的方法。此类测量对于研究肌钙蛋白水平(即收缩的基本单位)上的局部钙释放和机械反应非常重要,这对高血压,心房纤颤和心肌梗死等情况下的心脏功能障碍和心律不齐具有重要意义。在这里,我们描述了一种多模态二次谐波生成(SHG)和两个光子荧光(2PF)显微镜技术,该技术用于同时测量高空间和时间分辨率下的肌小节肌钙和收缩事件。该方法利用了SHG的无标记性质来对肉瘤成像以及SHG信号和钙指示剂激发的荧光信号的高空间共定位。该显微镜用于测量亚细胞微区中的钙火花和波动以及相关的收缩,从而导致亚细胞株的产生。我们预计这种新的成像工具将在研究机械应力诱发的心脏病中发挥重要作用。

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