A SYNERGY OF TECHNOLOGIES - COMBINING LASER MICROSURGERY WITH GREEN FLUORESCENT PROTEIN TAGGING

When focused through an objective lens with a high numerical aperture, nanosecond pulses of high-intensity green (532-nm) laser light can be used to selectively destroy any cellular component whose boundaries c an be defined by light microscopy. These components include, for examp le, chromosomes, spindle fibers, bundles of keratin, or actin filament s, mitochondria, vacuoles, and so forth. In addition, the definition o f poorly resolved components can be enhanced for selective destruction by tagging one or more of their constituent proteins with green fluor escence protein (GFP). As a example we show that the centrosome in liv ing PtK1 cells can be clearly defined, and then destroyed by green las er light, after transforming the cells with gamma-tubulin/GFP fusion p rotein. In some transformed cells it is even possible to target and se lectively destroy just one of the centrioles. (C) 1997 Wiley-Liss, Inc .