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Multiscale light-sheet for rapid imaging of cardiopulmonary system
Yichen Ding, … , René R. Sevag Packard, Tzung K. Hsiai
Yichen Ding, … , René R. Sevag Packard, Tzung K. Hsiai
Published August 23, 2018
Citation Information: JCI Insight. 2018;3(16):e121396. https://doi.org/10.1172/jci.insight.121396.
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Review

Multiscale light-sheet for rapid imaging of cardiopulmonary system

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Abstract

The ability to image tissue morphogenesis in real-time and in 3-dimensions (3-D) remains an optical challenge. The advent of light-sheet fluorescence microscopy (LSFM) has advanced developmental biology and tissue regeneration research. In this review, we introduce a LSFM system in which the illumination lens reshapes a thin light-sheet to rapidly scan across a sample of interest while the detection lens orthogonally collects the imaging data. This multiscale strategy provides deep-tissue penetration, high-spatiotemporal resolution, and minimal photobleaching and phototoxicity, allowing in vivo visualization of a variety of tissues and processes, ranging from developing hearts in live zebrafish embryos to ex vivo interrogation of the microarchitecture of optically cleared neonatal hearts. Here, we highlight multiple applications of LSFM and discuss several studies that have allowed better characterization of developmental and pathological processes in multiple models and tissues. These findings demonstrate the capacity of multiscale light-sheet imaging to uncover cardiovascular developmental and regenerative phenomena.

Authors

Yichen Ding, Jianguo Ma, Adam D. Langenbacher, Kyung In Baek, Juhyun Lee, Chih-Chiang Chang, Jeffrey J. Hsu, Rajan P. Kulkarni, John Belperio, Wei Shi, Sara Ranjbarvaziri, Reza Ardehali, Yin Tintut, Linda L. Demer, Jau-Nian Chen, Peng Fei, René R. Sevag Packard, Tzung K. Hsiai

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Figure 2

In vivo visualization of the developing hearts from the live zebrafish embryos and high-resolution imaging of chemotherapy-induced cardiac injury and regeneration.

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In vivo visualization of the developing hearts from the live zebrafish e...
(A–G) Light-sheet imaging of neural crest incorporation into the developing zebrafish heart tube (transgenic zebrafish line, Tg(-5sox10:GAL4,UAS:Cre)la2326Tg; Tg(UAS-E1b:NTR-mCherry)c264Tg; Tg(myl7:NLS-EGFP)chb2Tg). (A) Bright-field microscopic image of a zebrafish embryo 24 hpf. (B) The transgenic Tg(NC:NfsB-mCherry) zebrafish embryos express nuclear eGFP in the cardiomyocytes (green) and mCherry (red) in cells derived from the neural crest lineage at 36 hpf. (C–G) Colocalization of eGFP and mCherry from 26–30 hpf indicates the presence of cardiomyocytes of neural crest origin (yellow, arrows). (H–J) Dual-channel LSFM to capture the dynamic movement of atrioventricular (AV) valve leaflets from the transgenic Tg(fli1:GFP; cmlc2:mCherry) zebrafish embryos. An illustration of the AV valve leaflets in relation to the myocardium (cmlc:mCherry) and the endocardium (fli1:GFP) (H). AV valve leaflet closure (I) and opening (J) were captured at 100 fps. Myocardium (red) and endocardium (green) were concurrently acquired by the dual-channel imaging system. (K and L) Cardiac architecture following doxorubicin treatment and 3-D rendering of the adult zebrafish heart. A cross-section through the atrium, ventricle, and bulbus arteriosus demonstrates the two leaflets of the AV valve (red) and of the ventriculo-bulbar (VB) valve (orange) (K). Throughout the duration of the study, control hearts exhibited a preserved architecture in comparison with doxorubicin-treated groups at days 3, 30, and 60 (L). (Reproduced with permission from ref. 58). Scale bars: 50 μm (A–G); 10 μm (insets of C–G on the right side); 200 μm (I–L).

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