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Related Experiment Video

Updated: Jun 22, 2026

Multi-Photon Time Lapse Imaging to Visualize Development in Real-time: Visualization of Migrating Neural Crest Cells in Zebrafish Embryos
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Published on: August 9, 2017

In vivo developmental biology study using noninvasive multi-harmonic generation microscopy.

Shi-Wei Chu, Szu-Yu Chen, Tsung-Han Tsai

    Optics Express
    |May 28, 2009
    PubMed
    Summary

    This study introduces a noninvasive optical imaging method for observing intricate vertebrate embryo development. Higher harmonic generation microscopy allows clear, in vivo visualization of zebrafish embryos without prior treatment.

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    Last Updated: Jun 22, 2026

    Multi-Photon Time Lapse Imaging to Visualize Development in Real-time: Visualization of Migrating Neural Crest Cells in Zebrafish Embryos
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    Published on: August 9, 2017

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    Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

    Published on: September 30, 2021

    Area of Science:

    • Developmental Biology
    • Biomedical Optics
    • Microscopy

    Background:

    • Observing vertebrate embryonic development noninvasively at high resolution and deep penetration is challenging.
    • Existing methods often require sample treatment or lack simultaneous resolution and penetration capabilities.

    Purpose of the Study:

    • To develop and demonstrate a noninvasive optical microscopy technique for in vivo imaging of complex embryonic development.
    • To achieve sub-micrometer resolution with millimeter penetration depth in vertebrate embryos.

    Main Methods:

    • Utilized higher harmonic generation (second and third harmonics) as the intrinsic contrast mechanism.
    • Employed a 1230-nm near-infrared light source for deep penetration.
    • Leveraged the optical sectioning capability inherent in nonlinear harmonic generation.

    Main Results:

    • Successfully visualized morphological changes and developmental processes in zebrafish embryos in vivo.
    • Achieved clear imaging of early cell proliferation, gastrulation, and tissue formation.
    • Demonstrated noninvasive, label-free imaging without any treatment on live specimens.

    Conclusions:

    • Higher harmonic generation microscopy offers a powerful noninvasive tool for studying vertebrate embryonic development.
    • The technique overcomes limitations of resolution and penetration depth in live imaging.
    • Enables detailed in vivo observation of complex developmental events in zebrafish embryos.