Unit

UNIT 12.8 In Vivo Marking of Single Cells in Chick Embryos Using Photoactivation of GFP

  1. D. A. Stark,
  2. P. M. Kulesa

Published Online: 1 OCT 2005

DOI: 10.1002/0471143030.cb1208s28

Current Protocols in Cell Biology

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.

Author Information

  1. Stowers Institute for Medical Research, Kansas City, Missouri

Publication History

  1. Published Online: 1 OCT 2005
  2. Published Print: SEP 2005

Abstract

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.

Keywords:

  • photoactivation;
  • GFP;
  • chick;
  • embryo;
  • cell labeling;
  • lineage