PUBLICATION

Cell tracking using a photoconvertible fluorescent protein

Authors
Hatta, K., Tsujii, H., and Omura, T.
ID
ZDB-PUB-060921-8
Date
2006
Source
Nature Protocols   1(2): 960-967 (Journal)
Registered Authors
Hatta, Kohei
Keywords
none
MeSH Terms
  • Animals
  • Gene Expression Regulation
  • Luminescent Proteins/chemistry
  • Luminescent Proteins/metabolism*
  • Microscopy, Confocal/methods
  • Neurons/cytology
  • Neurons/metabolism
  • Zebrafish/embryology
PubMed
17406330 Full text @ Nat. Protoc.
Abstract
The tracking of cell fate, shape and migration is an essential component in the study of the development of multicellular organisms. Here we report a protocol that uses the protein Kaede, which is fluorescent green after synthesis but can be photoconverted red by violet or UV light. We have used Kaede along with confocal laser scanning microscopy to track labeled cells in a pattern of interest in zebrafish embryos. This technique allows the visualization of cell movements and the tracing of neuronal shapes. We provide illustrative examples of expression by mRNA injection, mosaic expression by DNA injection, and the creation of permanent transgenic fish with the UAS-Gal4 system to visualize morphogenetic processes such as neurulation, placode formation and navigation of early commissural axons in the hindbrain. The procedure can be adapted to other photoconvertible and reversible fluorescent molecules, including KikGR and Dronpa; these molecules can be used in combination with two-photon confocal microscopy to specifically highlight cells buried in tissues. The total time needed to carry out the protocol involving transient expression of Kaede by injection of mRNA or DNA, photoconversion and imaging is 2–8 d.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping