Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran

Joshua A. Clanton; Ilya A. Shestopalov; James K. Chen; Joshua T. Gamse

doi:10.3791/2672

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran

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Biology

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JoVE Journal Biology

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran

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07:35 min

April 28, 2011

DOI: 10.3791/2672-v

Joshua A. Clanton¹, Ilya A. Shestopalov², James K. Chen², Joshua T. Gamse¹

¹Department of Biological Sciences,Vanderbilt University, ²Department of Chemical and Systems Biology,Stanford University

This protocol delineates a way to label and trace the fate of small groups of cells zebrafish embryos using UV-uncaging of caged fluorescein, followed by whole mount immunolabeling to amplify the signal from the uncaged fluorescein.

The overall goal of this procedure is to lineage label cells in a zebrafish embryo. This is accomplished by first synthesizing caged fluorescein and injecting it into the zebrafish embryos. Embryos are then mounted in agarro so that the cells to be labeled are accessible.

Next, a UV laser is used to activate the caged fluorescein in selected cells of the embryo. Finally, embryos are harvested after they have developed to the desired stage and the labeled cells are detected in relation to markers of particular lineages. Ultimately, results can be obtained that show the fate of selected cells in the embryos through photo uncaging and immunofluorescent microscopy.

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