Amplification of the GFP signal localized in mitochondria of a transfected HeLa cell.
HeLa cell transfected with pShooter pCMV/myc/mito/GFP, then fixed and permeabilized. Green-fluorescent protein (GFP) localized in the mitochondria was labeled with anti-GFP mouse IgG2a (Cat. no. A11120) and detected with orange-fluorescent Alexa Fluor® 555 goat anti–mouse IgG (CAt. no. A21422), which colocalized with the dim GFP fluorescence. F-actin was labeled with green-fluorescent Alexa Fluor® 488 phalloidin (Cat. no. A12379), and the nucleus was stained with blue-fluorescent DAPI (Cat. no. D1306, D3571, D21490). The sample was mounted using ProLong® Gold antifade reagent (Cat. no. P36930). Some GFP fluorescence is retained in the mitochondria after fixation (top), but immunolabeling and detection greatly improve visualization (bottom).