Photoactivatable green fluorescent protein as a single-cell marker in living embryos

摘要: Selective marking of a single cell within an embryo is often difficult to perform with existing methods. Here, we report minimally invasive optical technique that uses 405-nm laser light photoactivate variant green fluorescent protein (PAGFP). Single cells and small groups (n < 10) are successfully marked, from region injected electroporated PAGFP, in both whole chick explants ovo. Photoactivated display normal migratory behaviors retain bright GFP signal for at least 24 hr when followed confocal time-lapse microscopy. We determined using low-magnification objective (∼ ×10) low power (∼1–10%) leads steady increase fluorescence photoactivated minimizes photobleaching. The utility PAGFP photoactivation was tested address specific question developmental biology. Specifically, asked whether neighboring emerge the hindbrain invade surrounding peripheral tissues maintain neighbor relationships while traveling destination sites. found some neural crest do not relationships, such two near tube may populate different branchial arches. ability optically or group follow individual living offers powerful, tool precise, vivo migration studies. Developmental Dynamics 233:983–992, 2005. © 2005 Wiley-Liss, Inc.