UNIT 12.8 In Vivo Marking of Single Cells in Chick Embryos Using Photoactivation of GFP
Published Online: 1 OCT 2005
Copyright © 2005 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Cell Biology
How to Cite
Stark, D. A. and Kulesa, P. M. 2005. In Vivo Marking of Single Cells in Chick Embryos Using Photoactivation of GFP. Current Protocols in Cell Biology. 28:12.8:12.8.1–12.8.11.
- Published Online: 1 OCT 2005
- Published Print: SEP 2005
Selective marking of a single cell within a living embryo is often difficult due to the inaccuracy and invasiveness of standard techniques. This unit describes a minimally invasive optical protocol that uses 405-nm laser light to photoactivate a variant of green fluorescent protein (PAGFP). This method takes advantage of the accessibility of the chick embryo to inject PAGFP into a region of interest and uses electroporation to deliver the construct into cells. This unit describes in detail how single and small groups of cells (n<10) that express PAGFP can be made visually distinguishable from the host population using the photoactivation process. Included is a means to maximize the fluorescence increase due to photoactivated GFP signal and to reduce photobleaching. Briefly outlined are previously developed chick culture and time-lapse imaging techniques to allow for the subsequent monitoring of photoactivated cell migratory behaviors. The technique has the potential to be a less-invasive, accurate tool for in vivo studies that involve following cell lineage and cell migration.
- cell labeling;