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Related Experiment Videos

Cell tracking using a photoconvertible fluorescent protein.

Kohei Hatta1, Hitomi Tsujii, Tomomi Omura

  • 1Laboratory for Vertebrate Body Plan, Center for Developmental Biology, RIKEN Kobe 2-2-3 Minatojima Minamimachi, Chuo-ku, Kobe 650-0047, Japan. khatta@cdb.riken.jp

Nature Protocols
|April 5, 2007
PubMed
Summary
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Researchers developed a new method using the Kaede protein and confocal microscopy to track cell development in zebrafish. This technique visualizes cell movement and neuronal shape, aiding the study of multicellular organism development.

Area of Science:

  • Developmental Biology
  • Cell Biology
  • Microscopy

Background:

  • Tracking cell fate, shape, and migration is crucial for understanding multicellular organism development.
  • Existing methods may have limitations in visualizing dynamic cellular processes.
  • Photoconvertible fluorescent proteins offer novel ways to label and track cells.

Purpose of the Study:

  • To present a protocol for tracking cell fate, shape, and migration using the photoconvertible protein Kaede.
  • To demonstrate the application of this technique in zebrafish embryos for visualizing developmental processes.
  • To adapt the protocol for other fluorescent molecules and advanced microscopy techniques.

Main Methods:

  • Utilizing the Kaede protein, which shifts from green to red fluorescence upon photoconversion with violet or UV light.

Related Experiment Videos

  • Employing confocal laser scanning microscopy to track labeled cells in zebrafish embryos.
  • Implementing mRNA injection, DNA injection for mosaic expression, and the UAS-Gal4 system for transgenic fish creation.

    • Main Results:

    • Successfully visualized cell movements and traced neuronal shapes in developing zebrafish.
    • Demonstrated the visualization of morphogenetic processes including neurulation, placode formation, and axon navigation.
    • Showcased the adaptability of the protocol for other fluorescent proteins (KikGR, Dronpa) and two-photon microscopy for deeper tissue imaging.

    Conclusions:

    • The Kaede-based photoconversion technique provides a powerful tool for studying cell dynamics during development.
    • This protocol enables detailed visualization of cellular processes in vivo.
    • The method is versatile and can be applied to various developmental contexts and advanced imaging setups.