Dynamic imaging of zebrafish heart with multi‐planar light sheet microscopy

Huang, Zengxin; Gu, Panchun; Kuang, Dengfeng; Ping, Mi; Feng, Xisheng

doi:10.1002/jbio.202000466

Cited by 5 publications

(1 citation statement)

References 31 publications

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“…Using a laser sheet to selectively illuminate the embryo along the detection focal plane, an entire section is captured at once with a camera, significantly reducing phototoxicity [46,47]. This approach has been successful at imaging primitive heart tube and looping stages in both in mouse [17,26,27,29] and zebrafish [46,48] live embryos.…”

Section: Fluorescence Microscopymentioning

confidence: 99%

Imaging Approaches and the Quantitative Analysis of Heart Development

Raiola

Sendra

Torres

2023

JCDD

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Heart morphogenesis is a complex and dynamic process that has captivated researchers for almost a century. This process involves three main stages, during which the heart undergoes growth and folding on itself to form its common chambered shape. However, imaging heart development presents significant challenges due to the rapid and dynamic changes in heart morphology. Researchers have used different model organisms and developed various imaging techniques to obtain high-resolution images of heart development. Advanced imaging techniques have allowed the integration of multiscale live imaging approaches with genetic labeling, enabling the quantitative analysis of cardiac morphogenesis. Here, we discuss the various imaging techniques used to obtain high-resolution images of whole-heart development. We also review the mathematical approaches used to quantify cardiac morphogenesis from 3D and 3D+time images and to model its dynamics at the tissue and cellular levels.

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Section: Fluorescence Microscopymentioning

confidence: 99%

Imaging Approaches and the Quantitative Analysis of Heart Development

Raiola

Sendra

Torres

2023

JCDD

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Linearly modulated multi-focal diffractive lens for multi-sheet excitation of flow driven samples in a light-sheet fluorescence microscope

Ghebjagh¹

2023

J. Eur. Opt. Society-Rapid Publ.

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Light sheet fluorescence microscope with single light sheet illumination enables rapid 3D imaging of living cells. In this paper we show the design, fabrication and characterization of a diffractive optical element producing several light sheets along a 45$^\circ$ inclined tube. The element, which is based on a multi-focal diffractive lens and a linear grating, generates five thin light sheets with equal intensities when combined with a refractive cylindrical lens. The generated uniform light sheets can be applied for the scanning of samples in tubes enabling flow-driven 3D imaging.

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Deep learning-based autofocus method enhances image quality in light-sheet fluorescence microscopy

Moatti

Zhang

et al. 2021

Biomed. Opt. Express

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Light-sheet fluorescence microscopy (LSFM) is a minimally invasive and high throughput imaging technique ideal for capturing large volumes of tissue with sub-cellular resolution. A fundamental requirement for LSFM is a seamless overlap of the light-sheet that excites a selective plane in the specimen, with the focal plane of the objective lens. However, spatial heterogeneity in the refractive index of the specimen often results in violation of this requirement when imaging deep in the tissue. To address this issue, autofocus methods are commonly used to refocus the focal plane of the objective-lens on the light-sheet. Yet, autofocus techniques are slow since they require capturing a stack of images and tend to fail in the presence of spherical aberrations that dominate volume imaging. To address these issues, we present a deep learning-based autofocus framework that can estimate the position of the objective-lens focal plane relative to the light-sheet, based on two defocused images. This approach outperforms or provides comparable results with the best traditional autofocus method on small and large image patches respectively. When the trained network is integrated with a custom-built LSFM, a certainty measure is used to further refine the network’s prediction. The network performance is demonstrated in real-time on cleared genetically labeled mouse forebrain and pig cochleae samples. Our study provides a framework that could improve light-sheet microscopy and its application toward imaging large 3D specimens with high spatial resolution.

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Dynamic imaging of zebrafish heart with multi‐planar light sheet microscopy

Cited by 5 publications

References 31 publications

Imaging Approaches and the Quantitative Analysis of Heart Development

Imaging Approaches and the Quantitative Analysis of Heart Development

Linearly modulated multi-focal diffractive lens for multi-sheet excitation of flow driven samples in a light-sheet fluorescence microscope

Deep learning-based autofocus method enhances image quality in light-sheet fluorescence microscopy

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