Xenopus Tadpole Craniocardiac Imaging Using Optical Coherence Tomography.
Xenopus Tadpole Craniocardiac Imaging Using Optical Coherence Tomography.
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DOI：
10.1101/pdb.prot105676
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发表时间：
2022-06-07
影响因子：
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通讯作者：
Khokha, Mustafa K
中科院分区：
文献类型：
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作者：
Deniz, Engin;Mis, Emily K;Khokha, Mustafa K
光学相干断层扫描（OCT）成像可以用来可视化的爪蟾模型系统中的颅心结构。OCT类似于超声波，利用光而不是声音来从样本反射的红外光的回波时间延迟创建灰度图像。OCT是一种高速、横截面、无标记成像模式，其可以以接近组织学细节的分辨率勾画动态体内形态。OCT成像可以真实的实时采集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内结构相关的血流模式。重要的是，OCT可以重复和有效地提供全面的，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。快速发展的非洲爪蟾模型结合快速OCT成像协议允许在短时间内识别特定的基因/致畸剂表型关系。功能丧失或获得实验可以在4-5天内进行评估，OCT成像仅需要5分钟/蝌蚪。因此，我们发现这种配对是一种有效的工作流程，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。
Optical coherence tomography (OCT) imaging can be used to visualize craniocardiac structures in the Xenopus 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 Xenopus 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.