Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)
Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)
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DOI：
10.3791/55002
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发表时间：
2016-12-01
影响因子：
1.2
通讯作者：
Zhou, Chao
中科院分区：
文献类型：
--
作者：
Men, Jing;Jerwick, Jason;Zhou, Chao
对小动物心跳的纵向研究有助于了解心脏发育过程中的结构和功能变化。光学相干显微镜（OCM）已被证明能够以高的空间分辨率和快速的成像速度对小动物心脏进行成像。高图像对比度和非侵入性特性使OCM成为进行纵向研究的理想选择，而无需组织解剖或染色。果蝇因其具有大量的人类疾病基因、分子机制和遗传途径与脊椎动物相似、生命周期短、培养成本低等特点，已被广泛用作心脏发育研究的模式生物。在这里，描述了实验协议的准备果蝇和光学成像的心跳与自定义OCM系统在整个生命周期的标本。按照本报告中提供的步骤，可以获得横向M模式和3D OCM图像，以进行果蝇心脏形态和功能的纵向研究。利用OCM的正、轴向截面图像、心率（HR）和心脏活动周期（CAP）直方图，结合M型图像构建的视频，直观地跟踪果蝇变态过程中心脏的活动，分析果蝇变态过程中心脏结构的变化，量化心脏动力学。由于果蝇与脊椎动物的遗传相似性，对果蝇心脏形态和动力学的纵向研究有助于揭示人类心脏疾病的起源。该方案将提供一种有效的方法来进行广泛的研究，以了解人类心脏疾病的机制。
Longitudinal study of the heartbeat in small animals contributes to understanding structural and functional changes during heart development. Optical coherence microscopy (OCM) has been demonstrated to be capable of imaging small animal hearts with high spatial resolution and ultrahigh imaging speed. The high image contrast and noninvasive properties make OCM ideal for performing longitudinal studies without requiring tissue dissections or staining. Drosophila has been widely used as a model organism in cardiac developmental studies due to its high number of orthologous human disease genes, its similarity of molecular mechanisms and genetic pathways with vertebrates, its short life cycle, and its low culture cost. Here, the experimental protocols are described for the preparation of Drosophila and optical imaging of the heartbeat with a custom OCM system throughout the life cycle of the specimen. By following the steps provided in this report, transverse M-mode and 3D OCM images can be acquired to conduct longitudinal studies of the Drosophila cardiac morphology and function. The en face and axial sectional OCM images and the heart rate (HR) and cardiac activity period (CAP) histograms, were also shown to analyze the heart structural changes and to quantify the heart dynamics during Drosophila metamorphosis, combined with the videos constructed with M-mode images to trace cardiac activity intuitively. Due to the genetic similarity between Drosophila and vertebrates, longitudinal study of heart morphology and dynamics in fruit flies could help reveal the origins of human heart diseases. The protocol here would provide an effective method to perform a wide range of studies to understand the mechanisms of cardiac diseases in humans.