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使用光学相干断层扫描进行蝌蚪颅心成像。

Tadpole Craniocardiac Imaging Using Optical Coherence Tomography.

机构信息

Pediatric Genomics Discovery Program, Department of Pediatrics,

Pediatric Genomics Discovery Program, Department of Pediatrics.

出版信息

Cold Spring Harb Protoc. 2022 Jun 7;2022(5):Pdb.prot105676. doi: 10.1101/pdb.prot105676.

Abstract

Optical coherence tomography (OCT) imaging can be used to visualize craniocardiac structures in the model system. OCT is analogous to ultrasound, utilizing light instead of sound to create a gray-scale image from the echo time delay of infrared light reflected from the specimen. OCT is a high-speed, cross-sectional, label-free imaging modality, which can outline dynamic in vivo morphology at resolutions approaching histological detail. OCT imaging can acquire 2D and 3D data in real time to assess cardiac and facial structures. Additionally, during cardiac imaging, Doppler imaging can be used to assess the blood flow pattern in relation to the intracardiac structures. Importantly, OCT can reproducibly and efficiently provide comprehensive, nondestructive in vivo cardiac and facial phenotyping. Tadpoles do not require preprocessing and thus can be further raised or analyzed after brief immobilization during imaging. The rapid development of the model combined with a rapid OCT imaging protocol allows the identification of specific gene/teratogen phenotype relationships in a short period of time. Loss- or gain-of-function experiments can be evaluated in 4-5 d, and OCT imaging only requires ∼5 min per tadpole. Thus, we find this pairing an efficient workflow for screening numerous candidate genes derived from human genomic studies to in-depth mechanistic studies.

摘要

光学相干断层扫描(OCT)成像可用于可视化模型系统中的颅心结构。OCT 类似于超声,利用光而不是声音,通过从样本反射的红外光的回波时间延迟来创建灰度图像。OCT 是一种高速、横截面、无标记的成像方式,可以在接近组织学细节的分辨率下描绘体内动态形态。OCT 成像可以实时获取 2D 和 3D 数据,以评估心脏和面部结构。此外,在心脏成像期间,多普勒成像可用于评估与心脏内结构相关的血流模式。重要的是,OCT 可以可重复且有效地提供全面、无损的体内心脏和面部表型。蝌蚪不需要预处理,因此可以在成像期间短暂固定后进一步饲养或分析。模型的快速发展与快速 OCT 成像方案相结合,使得在短时间内能够识别特定基因/致畸物表型关系。功能丧失或获得实验可以在 4-5 天内进行评估,并且 OCT 成像每个蝌蚪仅需要约 5 分钟。因此,我们发现这种组合是一种高效的工作流程,可用于筛选来自人类基因组研究的大量候选基因,以进行深入的机制研究。

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