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Cell Tracking Using Photoconvertible Proteins During Zebrafish Development

Cell Tracking Using Photoconvertible Proteins During Zebrafish Development

Published on: September 28, 2012

Cell tracking using photoconvertible proteins during zebrafish development.

Verónica A Lombardo¹, Anje Sporbert, Salim Abdelilah-Seyfried

¹Max Delbrück Center for Molecular Medicine.

Journal of Visualized Experiments : Jove

|October 12, 2012

Summary

This study introduces a new method for tracking cells in zebrafish embryos using photoconvertible fluorescent proteins (PCFPs). This technique allows researchers to follow specific cell populations throughout development, aiding in the study of embryogenesis.

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Area of Science:

Developmental Biology
Cell Biology
Genetics

Background:

Embryogenesis requires in vivo documentation of dynamic developmental changes.
Genetically-encoded fluorescent proteins facilitate the study of morphogenetic processes.
Photoactivatable and photoconvertible fluorescent proteins (PCFPs) enable tracking of specific protein subpopulations.

Purpose of the Study:

To develop and report a protocol for cell-specific photoconversion of PCFPs in living zebrafish embryos.
To enable tracking of photoconverted proteins in zebrafish at later developmental stages.
To facilitate the study of cell biological events underlying morphogenesis in a tissue-specific manner.

Main Methods:

Utilized photoconvertible fluorescent proteins (PCFPs) like Kaede, KikGR, Dendra, and EosFP.

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Employed light-induced (UV or blue light) photoconversion of the green chromophore to a red one.

Developed a protocol for cell-specific photoconversion in living zebrafish embryos for subsequent tracking.

Main Results:

Demonstrated successful cell-specific photoconversion of PCFPs in zebrafish embryos.
Enabled tracking of photoconverted cells and proteins through subsequent developmental stages.
Validated the methodology for tissue-specific analysis of cell behaviors during morphogenesis.

Conclusions:

The developed protocol provides a powerful tool for cell fate tracking in zebrafish.
This method enhances the study of cellular dynamics during embryogenesis and morphogenesis.
PCFPs offer advantages for in vivo imaging due to their ability to be tracked before and after photoconversion.