Tracking Cells in GFP-transgenic Zebrafish Using the Photoconvertible PSmOrange System

The overall goal of the PSmOrange Photo Conversion Technology is to track cells in the living animal during embryonic development and disease. This method can answer key questions in developmental biology such as uncovering the origin of cells which develop into tissues and organs. The main advantage of this technique is that it can be applied in GFP transgenic backgrounds.

This method can also be applied in different fields of investigation such as regeneration, tumor progression and invasion. After generating and embedding PSmOrange and GFP expressing embryos according to the text protocol, place the embryo under the confocal microscope and use the 20x air objective and 488 and 561 nanometer lasers to scan the specimen to identify an area to photoconvert. To detect the PSmOrange protein before photoconversion, use the 561 nanometer laser at 0.74 milliwatts of power measured at the focus plane above the objective and adjust the laser power as necessary.